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HELLO THERE, EVERYONE.

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WE HAVE A VERY FULL AGENDA TODAY

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SO I WILL TRY TO BE BRIEF MUCH

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WE HAVE AN EXCITING PROGRAM

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LINED UP FOR YOU IN THE FORM OF

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3 STELLAR YOUNG INVESTIGATORS

7
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WHO HAVE BEEN NOMINATE BIDE OUR

8
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SCIENTIFIC ADVISORY BOARD.

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BUT BEFORE I HAND THINGS OVER TO

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DR. MCINTOSH WHO WILL INTRODUCE

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THE FIRST SPEAKER, I WANT TO

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REMIND TO YOU PUT YOUR QUESTIONS

13
00:00:29,034 --> 00:00:34,472
IN THE CHAT BOX.

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OVER TO YOU EMPLOY

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>> THANK YOU, KAREN.

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GOOD AFTERNOON, EVERYONE.

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I'M DELIGHTED TO INTRODUCE OUR

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FIRST SPEAKER OF THE SESSION,

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TODAYING SPECIES AND D, INA

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REPLICATION IN THOSE SPECIES,

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FOLLOWING WHICH SHE DID A POST

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DOCTORAL FELLOWSHIP WITH

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DR. SAMUEL WILSON AT THE NIEHS,

24
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INVESTIGATE BASIC MOLECULAR

25
00:01:03,768 --> 00:01:04,936
MECHANISMS RELATED TO BASIC

26
00:01:04,936 --> 00:01:07,105
ECCISION REPAIR AND THEN IF 2019

27
00:01:07,105 --> 00:01:08,206
SHE WAS RECRUITED BY THE

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UNIVERSITY OF FLORIDA TO START

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AN INDEPENDENT CAREER AS A

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TENURED TRACK ASSISTANT

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PROFESSOR IN THE DEPARTMENT OF

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BIOCHEMIST RADIOY AND MOLECULAR

33
00:01:16,748 --> 00:01:16,981
BIOLOGY.

34
00:01:16,981 --> 00:01:20,685
IN HER INDEPENDENT POSITION, SHE

35
00:01:20,685 --> 00:01:22,354
CONTINUES TO STUDY THE PATHWAY

36
00:01:22,354 --> 00:01:24,622
PARTICULARLY LOOKING AT THE

37
00:01:24,622 --> 00:01:27,125
INTERPLAY BETWEEN THE DNA

38
00:01:27,125 --> 00:01:29,227
POLYMERASE BETA OR LIGASE 3 OR

39
00:01:29,227 --> 00:01:33,298
1A, NOW HER WORK HAS BEEN FUNDED

40
00:01:33,298 --> 00:01:36,634
BY A K99, R00 AS WELL AS MOST

41
00:01:36,634 --> 00:01:38,002
RECENTLY AN R35 EARLY STAGE

42
00:01:38,002 --> 00:01:39,671
INVESTIGATOR GRANT FROM THE

43
00:01:39,671 --> 00:01:44,743
NIGMS AND I'M LOOKING FORWARD TO

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00:01:44,743 --> 00:01:46,678
HEARING HER TELL US ABOUT HER

45
00:01:46,678 --> 00:01:50,315
RECENT WORK AND THE TITLE OF HER

46
00:01:50,315 --> 00:01:52,417
TALK IS NEXT STOPS ON THE ROAD

47
00:01:52,417 --> 00:01:54,018
TO DNA REPAIR EMPLOY NTHANK YOU

48
00:01:54,018 --> 00:01:55,620
SO MUCH FOR THE NICE

49
00:01:55,620 --> 00:01:57,122
INTRODUCTION AND HELLO EVERYONE

50
00:01:57,122 --> 00:01:59,491
EMPLOY I REALLY APPRECIATE THE

51
00:01:59,491 --> 00:02:03,027
OPPORTUNITY AND IT'S A GREAT

52
00:02:03,027 --> 00:02:04,195
PLEASURE TO PRESENT MY WORK TO

53
00:02:04,195 --> 00:02:04,696
THE GROUP.

54
00:02:04,696 --> 00:02:07,265
SO LET'S TART WITH THE

55
00:02:07,265 --> 00:02:08,700
DMA LIGASES, YOU KNOW VERY WELL,

56
00:02:08,700 --> 00:02:10,802
YOU KNOW DNA CAN BE GENERATED

57
00:02:10,802 --> 00:02:13,638
DURING NORMAL DECKER NA

58
00:02:13,638 --> 00:02:15,373
TRANSACTION OR DNA LESION BY

59
00:02:15,373 --> 00:02:25,884
EXCISION REPAIR AND THERE ARE

60
00:02:29,587 --> 00:02:36,294
POTENTIALLY--BY JOINING THE

61
00:02:36,294 --> 00:02:37,662
PRIME ENDS.

62
00:02:37,662 --> 00:02:40,365
THEREFORE D NA LIGASES ARE A

63
00:02:40,365 --> 00:02:42,667
FUNDAMENTAL ENZYME TO MAINTAIN

64
00:02:42,667 --> 00:02:43,368
THE STRUCTURAL

65
00:02:43,368 --> 00:02:44,035
ENTEGGIC--STRATEGICRITY OF THE

66
00:02:44,035 --> 00:02:45,136
HUMAN GENOME.

67
00:02:45,136 --> 00:02:47,338
PARTICULARLY THEY FINALAISE ALL

68
00:02:47,338 --> 00:02:49,574
DNA REPAIR PATHWAYS AND CLAIM

69
00:02:49,574 --> 00:02:54,179
IMPORTANT ROLE FOR THE

70
00:02:54,179 --> 00:03:00,218
[INDISCERNIBLE] LIGANDS WITH THE

71
00:03:00,218 --> 00:03:02,320
NUCLEACE REPLICATION.

72
00:03:02,320 --> 00:03:04,489
SO BASIC EXCISION REPAIR IS THE

73
00:03:04,489 --> 00:03:05,924
DNA REPAIR MECHANISM IN THE

74
00:03:05,924 --> 00:03:09,394
SIMILAR DEFENSE NEAR REMOVING

75
00:03:09,394 --> 00:03:11,029
SMALL AND OFFENSE

76
00:03:11,029 --> 00:03:13,398
NON[INDISCERNIBLE] BASAL LESIONS

77
00:03:13,398 --> 00:03:15,500
SUCH AS OXIDATION AND

78
00:03:15,500 --> 00:03:20,138
ONCUALATION PRODUCTS AS WELL AS

79
00:03:20,138 --> 00:03:21,406
SINGLE STRAND [INDISCERNIBLE].

80
00:03:21,406 --> 00:03:23,041
AND STEPS PROCEEDING IN AN

81
00:03:23,041 --> 00:03:25,143
ELDERLY MANNER AND HE RECEIVED A

82
00:03:25,143 --> 00:03:27,979
SHORT PATCH OR THE SINGLE BASIC

83
00:03:27,979 --> 00:03:29,581
EXCISION PAIR PATH, WHICH ARE

84
00:03:29,581 --> 00:03:32,317
RESPONSIBLE FOR THE REPAIR OF

85
00:03:32,317 --> 00:03:33,718
SINGLE DNA UNTIL [INDISCERNIBLE]

86
00:03:33,718 --> 00:03:36,054
AT THE DNA AT THE BEGINNING HERE

87
00:03:36,054 --> 00:03:38,623
AND THE BASIC EXCISION OF THE

88
00:03:38,623 --> 00:03:41,693
PATHWAY IS INITIATED BY SPECIFIC

89
00:03:41,693 --> 00:03:43,428
DNA GIVING--YOU COLSIS, MAYBE

90
00:03:43,428 --> 00:03:44,929
REMOVAL OF DECKER NA BASE

91
00:03:44,929 --> 00:03:47,599
SOLUTION, WHICH RESULTS IN THE

92
00:03:47,599 --> 00:03:48,500
FORMATION OF [INDISCERNIBLE] AND

93
00:03:48,500 --> 00:03:51,002
A PIECE SIDE AND THE CANNED WHAT

94
00:03:51,002 --> 00:03:54,706
IN A DOUBLE STRANDED DNA AND THE

95
00:03:54,706 --> 00:04:00,044
NEXT ENZYME, WITH THE APP AND 1

96
00:04:00,044 --> 00:04:01,880
AP1 [INDISCERNIBLE] BOUND AT

97
00:04:01,880 --> 00:04:05,149
LEAST TO THE RIBOPHOSPHATE BETA

98
00:04:05,149 --> 00:04:07,318
5 PRIME [INDISCERNIBLE].

99
00:04:07,318 --> 00:04:09,854
AND THEN DNA POLYMERASE IS A

100
00:04:09,854 --> 00:04:11,823
MAJOR CANNED WHAT POLYMERASE

101
00:04:11,823 --> 00:04:14,192
COME WITH A 2 CRITICAL FUNCTION

102
00:04:14,192 --> 00:04:17,695
FIRST THE POL REMOVES IT BY THE

103
00:04:17,695 --> 00:04:18,630
[INDISCERNIBLE] DNA LIGASE

104
00:04:18,630 --> 00:04:20,331
ACTIVITY AND THEN INCORPORATES

105
00:04:20,331 --> 00:04:21,533
THE [INDISCERNIBLE] TO FILL THE

106
00:04:21,533 --> 00:04:23,735
GAP DURING THE DNA SYNTHESIS TO

107
00:04:23,735 --> 00:04:25,136
REPAIR THE PATHWAY AND THIS

108
00:04:25,136 --> 00:04:26,104
CREATES [INDISCERNIBLE] IS A

109
00:04:26,104 --> 00:04:28,973
SUBSTRATE FOR THE NEXT STEP OF

110
00:04:28,973 --> 00:04:31,943
THE REPAIR PATHWAY WITH A 2

111
00:04:31,943 --> 00:04:35,380
PRIME AND 5 PRIME SEALED BY THE

112
00:04:35,380 --> 00:04:38,483
DID, NA LIGASE ALPHA AS A LAST

113
00:04:38,483 --> 00:04:39,884
TEP TO COMPLETE THE PATHWAY.

114
00:04:39,884 --> 00:04:42,720
AND THIS REPAIR PATHWAY

115
00:04:42,720 --> 00:04:45,623
COORDINATION IS FACILITATED BY

116
00:04:45,623 --> 00:04:46,324
MULTITELEPROTEIN, PROTEIN

117
00:04:46,324 --> 00:04:47,358
INTERACTION ON PROTEIN

118
00:04:47,358 --> 00:04:47,692
COMMUNICATION.

119
00:04:47,692 --> 00:04:50,862
IT HAS ALSO BEEN SHOWN IN

120
00:04:50,862 --> 00:04:52,163
[INDISCERNIBLE] IS POST DOC

121
00:04:52,163 --> 00:04:54,232
THERE IS A SUBSTRATE PRODUCT

122
00:04:54,232 --> 00:04:55,867
CHANNELING BETWEEN THE BR

123
00:04:55,867 --> 00:04:56,935
PROTEINS THAT INVOLVES THE

124
00:04:56,935 --> 00:04:58,469
HANDOFF OF REPAIRING

125
00:04:58,469 --> 00:04:59,370
[INDISCERNIBLE] FROM 1 STEP TO

126
00:04:59,370 --> 00:05:01,639
THE NEXT IN THAT SEQUENTIAL

127
00:05:01,639 --> 00:05:03,408
MANNER IN OTHER WORDS PRODUCT OF

128
00:05:03,408 --> 00:05:05,376
EACH STEP SERVE AS A SUBSTRATE

129
00:05:05,376 --> 00:05:07,412
FOR THE NEXT ENZYME IN THIS

130
00:05:07,412 --> 00:05:08,012
REPAIR PATHWAY.

131
00:05:08,012 --> 00:05:12,250
AND IT'S BEEN SHOWN ON THIS

132
00:05:12,250 --> 00:05:13,518
SUBSTRATE CHANNELING MECHANISM

133
00:05:13,518 --> 00:05:15,019
PREVENTS THE RELEASE AND

134
00:05:15,019 --> 00:05:16,621
ACCUMULATION OF THE SINGLE

135
00:05:16,621 --> 00:05:20,758
STRAND BREAK FOR THE CELL AND

136
00:05:20,758 --> 00:05:25,964
PERSIST OF THIS COULD TRIGGER

137
00:05:25,964 --> 00:05:28,633
SOME CELL CYCLE AND HOPEFUL

138
00:05:28,633 --> 00:05:29,300
NUCLEAR RECOMBINATION ACTIVITIES

139
00:05:29,300 --> 00:05:32,370
AND IN ADDITION TO THAT BR AND

140
00:05:32,370 --> 00:05:34,072
CO-BR PROTEINS I'M SHOWING HERE

141
00:05:34,072 --> 00:05:37,508
AND IT'S BEEN KNOWN UNDER SOME

142
00:05:37,508 --> 00:05:38,476
NONENZYMATIC SCAFFOLDING FACTORS

143
00:05:38,476 --> 00:05:39,510
SUCH AS [INDISCERNIBLE]

144
00:05:39,510 --> 00:05:43,881
ESSENTIAL FOR A PATIENT BR AND

145
00:05:43,881 --> 00:05:46,217
EXCIDE CC1 PLAY A ROLE IN THE

146
00:05:46,217 --> 00:05:47,251
PROMOTER REPAIR AND THE

147
00:05:47,251 --> 00:05:52,123
TREATMENT AND OF THE DNA DAMAGE

148
00:05:52,123 --> 00:05:53,858
AND PREVENTING THE FROM BEING

149
00:05:53,858 --> 00:05:55,927
RELEASED AND THE INDIVIDUAL

150
00:05:55,927 --> 00:05:57,962
STEPS HERE JUST STILL ON THE

151
00:05:57,962 --> 00:05:59,631
RULING TO HOLD THEM OUT TO THE

152
00:05:59,631 --> 00:06:03,635
BR COMPLEX, FOR THE REPAIR

153
00:06:03,635 --> 00:06:05,837
PATHWAY COORDINATION.

154
00:06:05,837 --> 00:06:08,072
AND THE DOWN STREAM STEEPS

155
00:06:08,072 --> 00:06:10,642
INVOLVING PATHWAY INVOLVING GUT

156
00:06:10,642 --> 00:06:12,510
FEELING AND POL-BETA AND

157
00:06:12,510 --> 00:06:15,013
DNALIGASE 1 AND 3 ALPHA AND

158
00:06:15,013 --> 00:06:16,848
EXCHANGEABLE BETWEEN THE

159
00:06:16,848 --> 00:06:17,215
[INDISCERNIBLE].

160
00:06:17,215 --> 00:06:20,084
AND REGARDING THE POLBETTA, IT'S

161
00:06:20,084 --> 00:06:23,388
THE SMALL [INDISCERNIBLE] DNA

162
00:06:23,388 --> 00:06:24,455
POLYMERASE, INCLUDING

163
00:06:24,455 --> 00:06:26,057
[INDISCERNIBLE] AND THE

164
00:06:26,057 --> 00:06:30,094
POLYMERASE DOMAIN FOR 3

165
00:06:30,094 --> 00:06:33,398
SUBDOMAINS, DNA BINDING, AND

166
00:06:33,398 --> 00:06:35,166
DNATPs, AND YOU CAN SEE IT IN

167
00:06:35,166 --> 00:06:38,569
THE STRUCTURAL MODEL OF THE

168
00:06:38,569 --> 00:06:38,803
PROTEIN.

169
00:06:38,803 --> 00:06:44,409
IT'S BEEN SHOWN BEAUTIFULLY BY

170
00:06:44,409 --> 00:06:46,177
[INDISCERNIBLE] OF HUMAN PERSON

171
00:06:46,177 --> 00:06:48,680
MUTATIONS CARRY THE PON, L-GENE

172
00:06:48,680 --> 00:06:51,416
AND IT CAN BE ASSOCIATED WITH

173
00:06:51,416 --> 00:06:53,518
VARIANTS IN MOUSE CELLS

174
00:06:53,518 --> 00:06:56,120
RESULLING IN PERMANENT CELL

175
00:06:56,120 --> 00:06:57,989
TRANSFORMATION [INDISCERNIBLE]

176
00:06:57,989 --> 00:07:00,324
TOXIC [INDISCERNIBLE].

177
00:07:00,324 --> 00:07:04,095
REGARDING THE DNA LIAISON CASES,

178
00:07:04,095 --> 00:07:07,965
THEY SHARE HIGHLY CONCERTED

179
00:07:07,965 --> 00:07:10,001
C-TERMINAL, FOR THE BINDING

180
00:07:10,001 --> 00:07:12,270
DOMAIN AND THE OTHER

181
00:07:12,270 --> 00:07:13,104
[INDISCERNIBLE] DOMAIN, END

182
00:07:13,104 --> 00:07:15,640
TERMINAL TO THE CAN THEA LYTIC,

183
00:07:15,640 --> 00:07:17,942
THERE'S A BINDING DOMAIN THAT

184
00:07:17,942 --> 00:07:21,145
BINDS MORE ROBUSTLY AND THE

185
00:07:21,145 --> 00:07:24,248
CATALYTIC CORE IS BINDED BY AN

186
00:07:24,248 --> 00:07:27,985
ACTIVE SEAT RESULTING IN THIS

187
00:07:27,985 --> 00:07:29,954
CORE AND CATALYTIC REAMINGION

188
00:07:29,954 --> 00:07:31,289
EXTENDS A CONFIRMATION IN THE

189
00:07:31,289 --> 00:07:34,992
ABSENCE OF DTHRKA, AND LACK

190
00:07:34,992 --> 00:07:36,260
CONFIRMATION CHANGE OF CURES

191
00:07:36,260 --> 00:07:38,362
WITHIN THE CORE THAT BINDS TO

192
00:07:38,362 --> 00:07:41,232
THE DNA BINDING DOMAIN TO FORM

193
00:07:41,232 --> 00:07:42,967
LIGASE PROTEIN CLAMP LIKE

194
00:07:42,967 --> 00:07:44,135
ARCHITECTURE WHICH CIRCLES THE

195
00:07:44,135 --> 00:07:46,104
[INDISCERNIBLE] WHICH YOU CAN

196
00:07:46,104 --> 00:07:48,773
SEE THIS STRUCTURE FOR BOTH DNA

197
00:07:48,773 --> 00:07:50,475
LIGASES.

198
00:07:50,475 --> 00:07:53,010
AND BOTH LIGASES HAVE A BLANKING

199
00:07:53,010 --> 00:07:54,445
UNRELATED END TERMINAL AND

200
00:07:54,445 --> 00:07:57,115
C-TERMINAL REGIONS AND

201
00:07:57,115 --> 00:07:57,815
DURING--THEY PLAY AN IMPORTANT

202
00:07:57,815 --> 00:08:00,318
PART IN THE SPECIFIC

203
00:08:00,318 --> 00:08:02,120
PROTEIN-PREEN TO INTERACTIONS

204
00:08:02,120 --> 00:08:05,623
WITH THE OTHER PROTEIN AND DNA

205
00:08:05,623 --> 00:08:11,395
REPAIR REPLICATION PROTEINS AND

206
00:08:11,395 --> 00:08:12,897
SUBSEQUENT LIGASES AND FUNCTION

207
00:08:12,897 --> 00:08:15,133
AND TO PARTICIPATE IN THE

208
00:08:15,133 --> 00:08:15,900
CELLULAR FUNK.

209
00:08:15,900 --> 00:08:17,135
TREAT END TERMINAL DOMAIN LIKE

210
00:08:17,135 --> 00:08:20,004
IS 1 THAT ACTS WITH THE PCA AND

211
00:08:20,004 --> 00:08:21,973
OTHER PROTEINS, SAME THING FOR

212
00:08:21,973 --> 00:08:24,809
INTERACTION BETWEEN LIGASE 3

213
00:08:24,809 --> 00:08:27,345
ALPHA AND P A RP1 EMPLOY AND

214
00:08:27,345 --> 00:08:30,014
IT'S BEEN KNOWN THAT THE SARS

215
00:08:30,014 --> 00:08:30,915
KRRK CWOBDERFUL--WONDERFUL MAKES

216
00:08:30,915 --> 00:08:33,217
A [INDISCERNIBLE] OFF LIGASE 1

217
00:08:33,217 --> 00:08:37,054
THAT OPERATING TO THE

218
00:08:37,054 --> 00:08:38,189
[INDISCERNIBLE] HISTORY

219
00:08:38,189 --> 00:08:38,823
SUBCELLULARLY.

220
00:08:38,823 --> 00:08:44,095
AND LIGATION REACTION IS ALSO

221
00:08:44,095 --> 00:08:44,796
HIGHLY CONSERVED.

222
00:08:44,796 --> 00:08:47,865
THIS USES A FRACTOR FOR THE

223
00:08:47,865 --> 00:08:50,601
REACTION INVOLVING 3 SUBSEQUENT

224
00:08:50,601 --> 00:08:54,505
CHEMICAL STEPS AND DURING STEP 1

225
00:08:54,505 --> 00:08:58,376
AND ON THE ATP LIGASE RESULTS IN

226
00:08:58,376 --> 00:09:03,147
THE FORMATION OF THE FIRST

227
00:09:03,147 --> 00:09:03,848
INTERMEDIATE LIGASE AMP.

228
00:09:03,848 --> 00:09:06,617
THIS IS LISTENED TO THE ACTIVE

229
00:09:06,617 --> 00:09:07,552
RESDUE RESIDING IN THE

230
00:09:07,552 --> 00:09:10,321
DOMINATION OF THE LIGASE AND YOU

231
00:09:10,321 --> 00:09:13,024
SEE THE [INDISCERNIBLE] 568 FOR

232
00:09:13,024 --> 00:09:13,958
LIGASE 1.

233
00:09:13,958 --> 00:09:15,726
AND DURING THAT STEP TO THE

234
00:09:15,726 --> 00:09:19,831
AMPOT LIGASE IS TRANSFERRED TO

235
00:09:19,831 --> 00:09:22,834
THE 5 PRIME, 5 PRIME END TO THE

236
00:09:22,834 --> 00:09:24,869
[INDISCERNIBLE] AND FORMING

237
00:09:24,869 --> 00:09:27,371
ANOTHER LIAISON GAGE REACTION

238
00:09:27,371 --> 00:09:28,639
TERM DNA, [INDISCERNIBLE] AND

239
00:09:28,639 --> 00:09:35,880
DURING THE LAST STEP 3 OFF THE

240
00:09:35,880 --> 00:09:37,081
LIGATION REACTION,

241
00:09:37,081 --> 00:09:38,482
[INDISCERNIBLE] ATTACKS THE

242
00:09:38,482 --> 00:09:40,918
[INDISCERNIBLE] DOWN STREAM TO

243
00:09:40,918 --> 00:09:42,653
JOIN THIS 3 PRIME WHICH IS

244
00:09:42,653 --> 00:09:45,189
COUPLED TO THE AMP RELEASE.

245
00:09:45,189 --> 00:09:47,925
AND THIS LIGATION REACTION

246
00:09:47,925 --> 00:09:51,729
REQUIRES THIS WAD WHAT.-

247
00:09:51,729 --> 00:09:52,196
-[INDISCERNIBLE].

248
00:09:52,196 --> 00:09:53,898
AND WE KNOW THAT THIS MAY FEEL

249
00:09:53,898 --> 00:09:55,666
IN THE DAMAGE OR MODIFIED 3

250
00:09:55,666 --> 00:09:57,301
PRIME EPPED RESULT NOTHING THE

251
00:09:57,301 --> 00:10:01,839
FORMATION OF LIGATION FAILURE

252
00:10:01,839 --> 00:10:03,241
PRODUCTS.

253
00:10:03,241 --> 00:10:05,376
AND THOSE LIGATION PRODUCT

254
00:10:05,376 --> 00:10:08,679
[INDISCERNIBLE] AND IT'S

255
00:10:08,679 --> 00:10:09,513
ACTUALLY LESIONS, AND THEREFORE

256
00:10:09,513 --> 00:10:11,816
IT'S FORMATION OF THE REPAIR OF

257
00:10:11,816 --> 00:10:16,287
[INDISCERNIBLE] ARE EXPECTED TO

258
00:10:16,287 --> 00:10:17,822
BE CRITICAL IN LIABILITY AND

259
00:10:17,822 --> 00:10:18,222
GENOME STABILITY.

260
00:10:18,222 --> 00:10:20,825
THERE ARE SOME ENZYMES AND

261
00:10:20,825 --> 00:10:23,895
PROCESSING TO FIGHT TO CARE OF

262
00:10:23,895 --> 00:10:26,364
THAT LIGATION PRODUCTS SUCH AS

263
00:10:26,364 --> 00:10:27,365
APT AND TOXIN.

264
00:10:27,365 --> 00:10:29,467
IT HAS BEEN SHOWN BEAUTIFULLY BY

265
00:10:29,467 --> 00:10:32,737
[INDISCERNIBLE] THAT THE LIGASE

266
00:10:32,737 --> 00:10:35,907
1 IS COVERED BY THE

267
00:10:35,907 --> 00:10:38,743
[INDISCERNIBLE] BY USING BINDING

268
00:10:38,743 --> 00:10:41,979
SITE AND SUBSTRATE TO RELEASE A

269
00:10:41,979 --> 00:10:42,680
[INDISCERNIBLE] DNA MIX.

270
00:10:42,680 --> 00:10:44,882
AS I SAID BEFORE, IN MY LAB

271
00:10:44,882 --> 00:10:46,384
STUDIES, DOWN STREAM STEPS ARE

272
00:10:46,384 --> 00:10:49,320
THOUGHT TO BE OUR

273
00:10:49,320 --> 00:10:52,723
[INDISCERNIBLE] BY GAP FILLING

274
00:10:52,723 --> 00:10:53,791
SUBSEQUENT [INDISCERNIBLE] AND

275
00:10:53,791 --> 00:10:57,161
POL BETA AND DNA LIGASES

276
00:10:57,161 --> 00:10:58,562
COORDINATE TO EFICIALLY PASS

277
00:10:58,562 --> 00:11:02,867
THROUGH THE INTERMEDIATE JUST

278
00:11:02,867 --> 00:11:04,735
LIKE INCOMING RUNEDDER VERSE US

279
00:11:04,735 --> 00:11:06,470
THE OTHER [INDISCERNIBLE].

280
00:11:06,470 --> 00:11:09,840
AND WE KNOW THAT THE DNA LIGASES

281
00:11:09,840 --> 00:11:12,410
WILL CURE SO IT INCORPORATES THE

282
00:11:12,410 --> 00:11:14,946
NUCLEOTIDINGS SUCH AS EGT AND

283
00:11:14,946 --> 00:11:16,480
CNA EXAMPLE YOU SEE HERE.

284
00:11:16,480 --> 00:11:18,816
IT CREATES A PERFECT SUBSTRATE

285
00:11:18,816 --> 00:11:21,085
FOR BOTH LIGASES WITH THE 3

286
00:11:21,085 --> 00:11:26,424
PRIME ENDS FOR EFFICIENT SEALING

287
00:11:26,424 --> 00:11:28,326
EMPLOY HOWEVER WE KNOW THAT

288
00:11:28,326 --> 00:11:33,531
UNDER HIGH STRESS CONDITIONS,

289
00:11:33,531 --> 00:11:34,932
[INDISCERNIBLE] POLBETTA CAN

290
00:11:34,932 --> 00:11:39,270
INCORPORATE THE MISMATCH, FOR

291
00:11:39,270 --> 00:11:44,408
THE OXIDIZED FORM OF IGTP A OR

292
00:11:44,408 --> 00:11:44,875
[INDISCERNIBLE].

293
00:11:44,875 --> 00:11:46,744
SIMILARLY IF THE POLBETTA IS

294
00:11:46,744 --> 00:11:47,378
ASSOCIATIONED AND ASSOCIATED

295
00:11:47,378 --> 00:11:52,650
MUSEUM TAIGS AS I MENTIONED

296
00:11:52,650 --> 00:11:54,385
BEFORE, IT HAS BEEN SHOWED THAT

297
00:11:54,385 --> 00:11:56,487
THE PROTEIN IS A REDUCED

298
00:11:56,487 --> 00:11:57,621
STABILITY AND/OR LACK OF GAP

299
00:11:57,621 --> 00:12:01,025
BUILDING AND THE QUESTIONS

300
00:12:01,025 --> 00:12:02,693
WHETHER THAT REPAIR IS COMING

301
00:12:02,693 --> 00:12:05,096
FROM THE POLBETTA WHERE IT CAN

302
00:12:05,096 --> 00:12:08,299
BE REACHED TO THE NEXT STEP IN

303
00:12:08,299 --> 00:12:09,433
LIGATION [INDISCERNIBLE] OR

304
00:12:09,433 --> 00:12:11,402
LIGASE 3 ALPHA.

305
00:12:11,402 --> 00:12:13,037
THAT'S WHERE WE HYPOTHESIZE THAT

306
00:12:13,037 --> 00:12:16,741
THE FAILURE IN THE DNA LIGASE

307
00:12:16,741 --> 00:12:19,210
MALFUNCTION SENDING THE PRODUCT

308
00:12:19,210 --> 00:12:21,278
CHANNELING IT DOWN THE STREAM

309
00:12:21,278 --> 00:12:23,314
FOR THE PATHWAY IT COULD BE

310
00:12:23,314 --> 00:12:24,382
SOURCE OF [INDISCERNIBLE]

311
00:12:24,382 --> 00:12:24,682
INSTABILITY.

312
00:12:24,682 --> 00:12:27,051
SO THIS IS PARTICULARLY FROM OUR

313
00:12:27,051 --> 00:12:28,085
STUDIES EMERGING THAT THE

314
00:12:28,085 --> 00:12:29,020
[INDISCERNIBLE] COULD CONTRIBUTE

315
00:12:29,020 --> 00:12:31,122
TO THE GENOMIC STABILITY TO

316
00:12:31,122 --> 00:12:33,290
NORMAL COORDINATION BETWEEN POL

317
00:12:33,290 --> 00:12:36,360
BETA AND LIGASES AND

318
00:12:36,360 --> 00:12:36,861
[INDISCERNIBLE].

319
00:12:36,861 --> 00:12:40,164
FOR EXAMPLE, IF THE POLBETTA

320
00:12:40,164 --> 00:12:42,767
CANNOT REMOVE THE DIT GROUP

321
00:12:42,767 --> 00:12:46,270
MOAGHT MAY ATTEMPT TO LIGATE

322
00:12:46,270 --> 00:12:49,006
[INDISCERNIBLE] ON THE P1 SIDE

323
00:12:49,006 --> 00:12:52,843
AND THE FORMATION OF THE

324
00:12:52,843 --> 00:12:55,012
REPAIRED [INDISCERNIBLE] WITH

325
00:12:55,012 --> 00:12:55,946
THE AN [INDISCERNIBLE] DIP GROUP

326
00:12:55,946 --> 00:12:59,116
HERE AND ANOTHER EXAMPLE,

327
00:12:59,116 --> 00:13:03,220
EXAMPLE INCORPORATION OF

328
00:13:03,220 --> 00:13:06,624
OXIDIZED GTPA, BY POLBETTA

329
00:13:06,624 --> 00:13:09,927
COMPRISES BOTH, AND RESULTS IN

330
00:13:09,927 --> 00:13:11,429
THE FORMATION, NOT OF ANOTHER

331
00:13:11,429 --> 00:13:14,432
[INDISCERNIBLE] BUT TO REPAIR

332
00:13:14,432 --> 00:13:16,000
INTERMEDIATE AND THEN 3 PRIME

333
00:13:16,000 --> 00:13:19,103
AND SORT OF ATOXIC GNP.

334
00:13:19,103 --> 00:13:22,006
I'VE SHOWN THERE IS SOME DETH

335
00:13:22,006 --> 00:13:24,141
PROCESSING ENZYME, CANNED WHAT,

336
00:13:24,141 --> 00:13:27,578
AND FAMILIES 1 CAN REMOVE THAT

337
00:13:27,578 --> 00:13:31,749
LIGATION FOR THE PRODUCTS FROM

338
00:13:31,749 --> 00:13:32,216
BOTH REPAIR.

339
00:13:32,216 --> 00:13:34,652
AND THIS IS JUST THAT THE

340
00:13:34,652 --> 00:13:36,387
DEFECTIVE SUBSTRATE PRODUCT

341
00:13:36,387 --> 00:13:37,922
CHANNELING FROM POL BETA TO THE

342
00:13:37,922 --> 00:13:39,924
DID, NA LIGASE CAN LEAD TO THE

343
00:13:39,924 --> 00:13:42,893
INTERACTIONS IN THE BR PATHWAY

344
00:13:42,893 --> 00:13:45,096
ACCORD NATION AND RESULTING IN

345
00:13:45,096 --> 00:13:51,168
THE EVENTS AND DNA INTERMEDUOUS 

346
00:13:51,168 --> 00:13:53,337
EVER SINCE THIS, WE CONTRIBUTE

347
00:13:53,337 --> 00:13:56,107
TO FIND A KEY MOLECULAR

348
00:13:56,107 --> 00:13:57,208
DETERMINANT THAT INSURE THE

349
00:13:57,208 --> 00:14:00,444
COORDINATION OR RESULTS IN THE

350
00:14:00,444 --> 00:14:02,012
HANDLE FROM POL-BETA TO DNA

351
00:14:02,012 --> 00:14:02,246
LIGASE.

352
00:14:02,246 --> 00:14:05,649
FOR EXAMPLE, WE HAVE SHOWN THAT

353
00:14:05,649 --> 00:14:07,618
HOW THE IDENTICAL MISMATCH

354
00:14:07,618 --> 00:14:11,889
INCORPORATED BY POL BETA COULD

355
00:14:11,889 --> 00:14:15,292
ASHING THAT EFFICIENT OF THE

356
00:14:15,292 --> 00:14:16,427
FINAL STEPS HERE.

357
00:14:16,427 --> 00:14:26,937
OUR FINDINGS SUGGEST THAT THE

358
00:14:29,006 --> 00:14:29,640
PRODUCT MATCH [INDISCERNIBLE]

359
00:14:29,640 --> 00:14:32,076
BUT HOWEVER, FOR THE PARTICULAR

360
00:14:32,076 --> 00:14:34,879
MISMATCH, THE GTP INSERTION

361
00:14:34,879 --> 00:14:36,614
POSITION C, NOW MISMATCH

362
00:14:36,614 --> 00:14:37,481
INSERTION ARE COMPATIBLE WITH

363
00:14:37,481 --> 00:14:39,517
THE PROCESS LEADING TO THE

364
00:14:39,517 --> 00:14:42,253
LIAISON GAGE AFTER POLBETTA GT

365
00:14:42,253 --> 00:14:44,555
AND CANNED WHAT CONSORTIUM

366
00:14:44,555 --> 00:14:45,189
PRODUCTS.

367
00:14:45,189 --> 00:14:45,823
[INDISCERNIBLE] ALL OTHER

368
00:14:45,823 --> 00:14:49,059
MISMATCH WE RECEIVE THE DATP C,

369
00:14:49,059 --> 00:14:51,595
THAT INSERTION AND THAT PRODUCT

370
00:14:51,595 --> 00:14:56,901
CANNOT BE CHANNELED TO THE NEXT

371
00:14:56,901 --> 00:14:58,869
LIGATION SET, WHICH COULD

372
00:14:58,869 --> 00:15:02,706
PROVIDE AN OPPORTUNITY FOR

373
00:15:02,706 --> 00:15:03,707
OPPORTUNITY PROCESSING ENZYME

374
00:15:03,707 --> 00:15:06,377
AND COME IN TO PROOF READ THAT

375
00:15:06,377 --> 00:15:08,012
AND MISMATCH AS WE'VE SHOWN FOR

376
00:15:08,012 --> 00:15:10,080
P1 AND BEAUTIFULLY SHOWN FOR THE

377
00:15:10,080 --> 00:15:13,150
STRUCTURE FOR THEM LIFE AND IT

378
00:15:13,150 --> 00:15:15,186
WAS THAT GO BACK TO THE PAINT

379
00:15:15,186 --> 00:15:18,088
AND GO BACK TO THE SITES FOR AN

380
00:15:18,088 --> 00:15:21,725
ATTEMPT FOR POL BETA

381
00:15:21,725 --> 00:15:22,826
INCORPORATION.

382
00:15:22,826 --> 00:15:27,765
AND WE ALSO DEMONSTRATE THAT THE

383
00:15:27,765 --> 00:15:29,500
LIGASE IS COMPROMISED BY

384
00:15:29,500 --> 00:15:31,802
[INDISCERNIBLE] IN ALL POSSIBLE

385
00:15:31,802 --> 00:15:33,871
CANNONICLE BASE PAIRS AND IT'S

386
00:15:33,871 --> 00:15:36,307
IN THE INTERMEDIATE LIGASE AND

387
00:15:36,307 --> 00:15:38,943
THE LIGATION AND DNA SUBSTRATE

388
00:15:38,943 --> 00:15:41,111
WITH THE 3 PRIME [INDISCERNIBLE]

389
00:15:41,111 --> 00:15:43,714
AND ALL THE 3 PRIME INSERTED

390
00:15:43,714 --> 00:15:45,649
CORRECT THE BEST PART AND

391
00:15:45,649 --> 00:15:46,917
[INDISCERNIBLE] AND THE MISMATCH

392
00:15:46,917 --> 00:15:49,420
AND FOR EXAMPLE, GT AND AC, YOU

393
00:15:49,420 --> 00:15:50,187
SEE HERE.

394
00:15:50,187 --> 00:15:52,890
AND THE RESULTS SHOW THAT THE

395
00:15:52,890 --> 00:15:54,758
[INDISCERNIBLE] CAN BE EFFICIENT

396
00:15:54,758 --> 00:15:56,393
LIGATED WHERE WE WERE ABLE TO

397
00:15:56,393 --> 00:15:59,029
SEE THE SAME EFFICIENCY FOR THE

398
00:15:59,029 --> 00:16:00,864
PARTICULAR SUBSTRATE INCLUDING

399
00:16:00,864 --> 00:16:01,232
AC MUCH.

400
00:16:01,232 --> 00:16:05,502
AND ACTUALLY THIS HAS BEEN

401
00:16:05,502 --> 00:16:06,637
SIMILARLY THE OBSERVED DISTINCT

402
00:16:06,637 --> 00:16:08,872
ABILITY OF THE LIGASE 1 VERSUS

403
00:16:08,872 --> 00:16:10,608
LIGASE 3 ALPHA TO LIKE THE

404
00:16:10,608 --> 00:16:13,944
NUCLEAR REPAIR IN TIME WITH THE

405
00:16:13,944 --> 00:16:15,746
PREINSERTED MISMATCH DEPENDING

406
00:16:15,746 --> 00:16:17,281
ON THE CHEMICAL CHARACTERISTIC

407
00:16:17,281 --> 00:16:19,984
OF ALL 12 POSSIBLE NONCANNONICLE

408
00:16:19,984 --> 00:16:22,152
BEST PAIRS BETWEEN THE PRIMARY

409
00:16:22,152 --> 00:16:24,822
TERMINALS AND THE TEMPLATE BASE,

410
00:16:24,822 --> 00:16:29,360
THEY ARE--THEY MIMIC THE

411
00:16:29,360 --> 00:16:30,094
BIOLOGICALLY MYCK SUBSTRATE

412
00:16:30,094 --> 00:16:35,899
BEING BE FORMED DUE TO THE

413
00:16:35,899 --> 00:16:36,467
MISMATCHED [INDISCERNIBLE]

414
00:16:36,467 --> 00:16:38,836
INCORPORATIONS. 

415
00:16:38,836 --> 00:16:41,372
AND OUR STRUCTURES APPLIED AND

416
00:16:41,372 --> 00:16:42,673
ALSO REVEALED THAT 1 ACTIVE

417
00:16:42,673 --> 00:16:45,209
PROCESS WE DETERMINED WITH THE

418
00:16:45,209 --> 00:16:48,512
MISMATCH DISTINCTLY, WE WERE

419
00:16:48,512 --> 00:16:50,614
ABLE TO SO THAT THE LIAISON

420
00:16:50,614 --> 00:16:52,349
ANYPLACE CAN ACCOMMODATE THE

421
00:16:52,349 --> 00:16:54,084
MISMATCH IN A CONFIRMATION

422
00:16:54,084 --> 00:16:56,620
DURING STEP 2 AFTER LIGATION

423
00:16:56,620 --> 00:17:02,393
REACTION AND THEN

424
00:17:02,393 --> 00:17:03,460
[INDISCERNIBLE] NICK DNA, AND

425
00:17:03,460 --> 00:17:06,530
THEY ALSO CAPTURE THE LIGASE 1

426
00:17:06,530 --> 00:17:09,633
AT THE SAME STEPS AS STEP WITH

427
00:17:09,633 --> 00:17:12,102
THE NICK DNA CONTAINING THE

428
00:17:12,102 --> 00:17:12,836
CANNED WHAT.

429
00:17:12,836 --> 00:17:16,573
HOWEVER, WE FOUND THAT THE

430
00:17:16,573 --> 00:17:17,775
ACTIVE SITES STATES ARE

431
00:17:17,775 --> 00:17:18,842
VENTILATED DURING THE INITIAL

432
00:17:18,842 --> 00:17:22,379
STEP 1 OF THE LIGATION REACTION,

433
00:17:22,379 --> 00:17:25,316
WHILE ENGAGING WITH THE NICK DNA

434
00:17:25,316 --> 00:17:27,051
CONTAINING [INDISCERNIBLE], AND

435
00:17:27,051 --> 00:17:30,821
RECEIVED THE AMP, AND THE

436
00:17:30,821 --> 00:17:34,525
RESIDUE, WITH THE FIRST LIGATION

437
00:17:34,525 --> 00:17:34,758
STEP.

438
00:17:34,758 --> 00:17:38,162
AND THEN THIS STUDY CAN FORWARD

439
00:17:38,162 --> 00:17:39,330
LIGASE [INDISCERNIBLE] FOR

440
00:17:39,330 --> 00:17:42,766
LIGASE 1, THAT FAVORS TO GT OR

441
00:17:42,766 --> 00:17:46,837
DETER THE AC AND MISMATCH, THE

442
00:17:46,837 --> 00:17:50,808
LIGATION OF THE BASIS

443
00:17:50,808 --> 00:17:51,875
SUBSTITUTION ERROR INCORPORATED

444
00:17:51,875 --> 00:17:53,877
AND INCORPORATED BY THE DNA

445
00:17:53,877 --> 00:17:58,282
POLYMERASES, AND WE ALSO TEST

446
00:17:58,282 --> 00:17:59,650
THE LIGATIONENTIOUS FICIENT

447
00:17:59,650 --> 00:18:01,952
STUFF, FOR THE SUBSTRATE WITH

448
00:18:01,952 --> 00:18:05,689
THE INSERTED 3 PRIMER

449
00:18:05,689 --> 00:18:06,757
[INDISCERNIBLE] SIDES OUR

450
00:18:06,757 --> 00:18:08,325
RESULTS DEMONSTRATE THE SEALING

451
00:18:08,325 --> 00:18:13,097
OF THAT TO RECEIVE THE 3--FOR

452
00:18:13,097 --> 00:18:16,333
TREATING THE 3 PRIME RIBOAT,

453
00:18:16,333 --> 00:18:19,269
RIBOTC AND RIBOGC, WHICH WAS

454
00:18:19,269 --> 00:18:23,207
COMPARABLE WITH THE NORMAL

455
00:18:23,207 --> 00:18:26,710
COUNTER FOR THE [INDISCERNIBLE]

456
00:18:26,710 --> 00:18:27,010
NUCLEOTIDES.

457
00:18:27,010 --> 00:18:29,913
IS THIS ACTUALLY FOR ALL OTHER

458
00:18:29,913 --> 00:18:33,150
12S MISMATCH 20 FOR RIBOGA AND

459
00:18:33,150 --> 00:18:34,885
DEMONSTRATING THE

460
00:18:34,885 --> 00:18:36,086
[INDISCERNIBLE] DISCRIMINATION

461
00:18:36,086 --> 00:18:37,521
AGAINST RIBONUCLEOTIDES

462
00:18:37,521 --> 00:18:40,391
TREATMENT TO THE REPAIR, THAT

463
00:18:40,391 --> 00:18:41,658
COULD [INDISCERNIBLE] INSERTION

464
00:18:41,658 --> 00:18:43,660
PRODUCTS AT THE DNA POLYMERASES

465
00:18:43,660 --> 00:18:46,430
AND YOU ALSO SOLD STRUCTURAL

466
00:18:46,430 --> 00:18:48,532
[INDISCERNIBLE] DNA COMPLEX,

467
00:18:48,532 --> 00:18:49,533
CONTAINING THAT SIMILAR

468
00:18:49,533 --> 00:18:49,967
RIBO[INDISCERNIBLE].

469
00:18:49,967 --> 00:18:53,837
AND HERE TO SEE THE STRUCTURE

470
00:18:53,837 --> 00:18:56,006
FOR RIBOAT TO CAPTURE THE ACTIVE

471
00:18:56,006 --> 00:18:58,642
SITE AT THE LAST 3 WHICH IS THE

472
00:18:58,642 --> 00:19:02,312
LAST LIGATION STEP OF--LAST STEP

473
00:19:02,312 --> 00:19:03,847
OF LIGATION REACTION, AND 3

474
00:19:03,847 --> 00:19:07,251
PRIME AND 5 PRIME AND RESPOND.

475
00:19:07,251 --> 00:19:10,988
IN STEP 3 STRUCTURE, SHOWS AT

476
00:19:10,988 --> 00:19:12,556
THE AMPs COMPLEX SUGGIESTING

477
00:19:12,556 --> 00:19:19,062
THAT THE AMPs RELEASE, AND

478
00:19:19,062 --> 00:19:20,431
THIS IS SUGGESTING THAT

479
00:19:20,431 --> 00:19:23,367
[INDISCERNIBLE] FROM BETWEEN THE

480
00:19:23,367 --> 00:19:27,371
3 PRIME AND 5 PRIME DNA FOR THIS

481
00:19:27,371 --> 00:19:27,871
TO CONTINUE.

482
00:19:27,871 --> 00:19:31,175
AND I THINK THAT'S ALL FOR

483
00:19:31,175 --> 00:19:38,682
TODAY, I WOULD LIKE TO TURN TO

484
00:19:38,682 --> 00:19:40,851
MY MEMBER [INDISCERNIBLE] AND

485
00:19:40,851 --> 00:19:43,487
ALSO THE FUNDING INTERNAL AND

486
00:19:43,487 --> 00:19:44,021
EXTERNAL FUNDING.

487
00:19:44,021 --> 00:19:46,457
HAPPY TO TAKE QUESTIONS. 

488
00:19:46,457 --> 00:19:47,524
NTHANK YOU.

489
00:19:47,524 --> 00:19:48,559
>> LET ME READ THE FIRST

490
00:19:48,559 --> 00:19:51,895
QUESTION I SEE HERE, IT'S FROM

491
00:19:51,895 --> 00:19:52,362
[INDISCERNIBLE].

492
00:19:52,362 --> 00:19:58,836
HAVE YOU BEEN ABLE TO SEE IF

493
00:19:58,836 --> 00:20:00,204
KNOWN B-DNA STRUCTURES FORM IN

494
00:20:00,204 --> 00:20:01,605
THE PROCESS AND IF THEY'RE

495
00:20:01,605 --> 00:20:03,474
INVOLVED IN ANY WAY OF THE NICK

496
00:20:03,474 --> 00:20:03,640
IN.

497
00:20:03,640 --> 00:20:05,209
>> NO, WE HAVE NOT.

498
00:20:05,209 --> 00:20:12,916
WE HAVE NOT SEEN THAT COME FROM

499
00:20:12,916 --> 00:20:13,517
THE STRUCTURES.

500
00:20:13,517 --> 00:20:14,384
>> SO WHILE PEOPLE ARE
THINKING

501
00:20:14,384 --> 00:20:15,819
OF ARK DITIONAL QUESTIONS,

502
00:20:15,819 --> 00:20:19,189
PERHAPS I COULD ASK 1.

503
00:20:19,189 --> 00:20:20,724
SO YOU MENTIONED THERE ARE THIS

504
00:20:20,724 --> 00:20:24,361
CANCERS YOU CAN HAVE VARIANTS OF

505
00:20:24,361 --> 00:20:28,632
POL-BETA, RIGHT IN SO ARE

506
00:20:28,632 --> 00:20:30,167
THOSE--ARE THOSE VARIANTS

507
00:20:30,167 --> 00:20:31,468
POL BETA MORE MUTE O GENIC AND

508
00:20:31,468 --> 00:20:37,774
IS THERE A CORRELATION BETWEEN

509
00:20:37,774 --> 00:20:38,675
THE PARTICULAR MUTATION

510
00:20:38,675 --> 00:20:40,844
[INDISCERNIBLE] IN THE ENZYME

511
00:20:40,844 --> 00:20:43,680
AND ANY PARTICULAR TYPE OF

512
00:20:43,680 --> 00:20:46,483
MISMATCH THAT CAN HAPPEN IN.

513
00:20:46,483 --> 00:20:47,317
>> RIGHT.

514
00:20:47,317 --> 00:20:48,585
SO THE PARAMETERS IS A

515
00:20:48,585 --> 00:20:49,887
PREFERENCE BETWEEN A PARTICULAR

516
00:20:49,887 --> 00:20:51,722
TYPE OF MISMATCH HAS BEEN SHOWN

517
00:20:51,722 --> 00:20:54,858
BEFORE, I DON'T REMEMBER THAT

518
00:20:54,858 --> 00:20:56,026
PARTICULAR [INDISCERNIBLE] IN MY

519
00:20:56,026 --> 00:20:57,227
MIND SO IT'S REALLY A LOT BUT

520
00:20:57,227 --> 00:20:59,763
SOME OF THE [INDISCERNIBLE] IS A

521
00:20:59,763 --> 00:21:00,697
PREFERENCE TO INSERT A

522
00:21:00,697 --> 00:21:03,200
PARTICULAR TYPE OF MISMATCH, FOR

523
00:21:03,200 --> 00:21:04,568
SOME REASON [INDISCERNIBLE] DOES

524
00:21:04,568 --> 00:21:06,370
NOT DO GAP FILLING AT ALL, FOR

525
00:21:06,370 --> 00:21:08,405
SOME REASON, CANNED WHAT IS SOME

526
00:21:08,405 --> 00:21:18,615
OTHER FORM OF THE

527
00:21:18,615 --> 00:21:19,616
[INDISCERNIBLE].

528
00:21:19,616 --> 00:21:19,917
NTHANK YOU.

529
00:21:19,917 --> 00:21:22,953
I SEE A QUESTION FROM DEAN ROTH

530
00:21:22,953 --> 00:21:25,222
WHAT DETERMINES THE LIGASE 1 OR

531
00:21:25,222 --> 00:21:25,989
3 ALPHA?

532
00:21:25,989 --> 00:21:28,692
>> WHAT DETERPS THE
CHOICE?

533
00:21:28,692 --> 00:21:32,930
>> SO SO WHAT DETERMINES
THE

534
00:21:32,930 --> 00:21:33,163
CHOICE.

535
00:21:33,163 --> 00:21:33,931
>> THEY'RE EXCHANGEABLE.

536
00:21:33,931 --> 00:21:35,365
THERE ARE 2, THAT IS WHAT I

537
00:21:35,365 --> 00:21:37,334
INTRODUCED TODAY AND THERE'S

538
00:21:37,334 --> 00:21:40,203
ANOTHER SUBTYPE THAT SHOWS A

539
00:21:40,203 --> 00:21:41,438
LONG PATCH [INDISCERNIBLE].

540
00:21:41,438 --> 00:21:44,341
STUDIES HAVE BEEN SHOWN THAT

541
00:21:44,341 --> 00:21:46,710
LIGASE 1 INVOLVING MOSTLY THE

542
00:21:46,710 --> 00:21:48,278
LONG [INDISCERNIBLE] SO INCLUDES

543
00:21:48,278 --> 00:21:50,747
OTHER REPAIR PROTEINS LIKE PCA

544
00:21:50,747 --> 00:21:52,149
AND REPLICATED POLYMERASES,

545
00:21:52,149 --> 00:21:54,685
LIGASE HISTORY, AND LIGASE AND

546
00:21:54,685 --> 00:21:56,987
FINAL ICING THE SHORT PATH AND

547
00:21:56,987 --> 00:21:58,622
THE VR, BUT EXCHANGEABILITY AND

548
00:21:58,622 --> 00:22:01,191
THEN IN CASE OF DEFICIENCY IN

549
00:22:01,191 --> 00:22:04,795
LIGASE 1, LIGASE 3 ALPHA CAN

550
00:22:04,795 --> 00:22:07,464
COMPLEMENT THE NICK SEALING

551
00:22:07,464 --> 00:22:07,931
[INDISCERNIBLE].

552
00:22:07,931 --> 00:22:08,231
>> GREAT.

553
00:22:08,231 --> 00:22:09,733
THANK YOU SO MUCH.

554
00:22:09,733 --> 00:22:11,401
I THINK IN THE INTEREST OF TIME.

555
00:22:11,401 --> 00:22:15,505
WE WILL MOVE ON AND I WILL HAND

556
00:22:15,505 --> 00:22:18,508
OVER TO PATTY.

557
00:22:18,508 --> 00:22:19,376
>> HI, IT'S GREAT TO SEE

558
00:22:19,376 --> 00:22:20,644
EVERYBODY, I HAVE A SPECIAL

559
00:22:20,644 --> 00:22:22,179
GUEST WITH ME THIS YEAR, HE'S

560
00:22:22,179 --> 00:22:23,413
VISITING US AND HE GETS TO SEE

561
00:22:23,413 --> 00:22:26,516
THE TALKS TODAY AS WELL BECAUSE

562
00:22:26,516 --> 00:22:28,785
NORMALLY HE'S--IT'S IN THE

563
00:22:28,785 --> 00:22:31,355
MIDDLE OF THE NIGHT FOR HIM.

564
00:22:31,355 --> 00:22:33,223
SO WELCOME ORLANDO.

565
00:22:33,223 --> 00:22:33,824
>> HI, EVERYBODY.

566
00:22:33,824 --> 00:22:35,125
>> SO ON MY DISTRIBUTION,
IT'S

567
00:22:35,125 --> 00:22:40,263
MY GREAT PLEASURE TO INTRODUCE

568
00:22:40,263 --> 00:22:41,465
DR. ELISE, [INDISCERNIBLE], SHE

569
00:22:41,465 --> 00:22:44,334
RECEIVED HER Ph.D. WHERE SHE

570
00:22:44,334 --> 00:22:48,705
BEGAN HER ADVENTURES IN THE

571
00:22:48,705 --> 00:22:53,844
PARTNERSHIP 1 ENZYMES AND FROM

572
00:22:53,844 --> 00:22:55,145
THERE SHE MOVED TO UNIVERSITY OF

573
00:22:55,145 --> 00:22:56,647
PITTSBURGH AND THEN I WAS

574
00:22:56,647 --> 00:22:58,181
FORTUNATE TO RECRUIT HER TO MY

575
00:22:58,181 --> 00:23:01,051
LAB WHERE SHE DID A SECOND POST

576
00:23:01,051 --> 00:23:01,251
DOCK.

577
00:23:01,251 --> 00:23:02,619
SHE WAS INKRE TINSIBLELY

578
00:23:02,619 --> 00:23:04,955
SUCCESSFUL, AND EARNED A K99 AND

579
00:23:04,955 --> 00:23:06,490
THEN WENT TO THOMAS JEFFERSON

580
00:23:06,490 --> 00:23:09,559
UNIVERSITY WHERE SHE STARTED HER

581
00:23:09,559 --> 00:23:13,397
LAB AND SHE WAS AWARDED AN R35,

582
00:23:13,397 --> 00:23:15,465
SO SHE HIT THE GROUND RUNNING

583
00:23:15,465 --> 00:23:17,200
AND THEN WE WERE VERY FORTUNATE

584
00:23:17,200 --> 00:23:18,935
TO BE ABLE TO RECRUIT HER BACK

585
00:23:18,935 --> 00:23:19,670
TO THE UNIVERSITY OF PITTSBURGH

586
00:23:19,670 --> 00:23:21,638
WHERE SHE IS AN ASSISTA ANT

587
00:23:21,638 --> 00:23:24,174
PROCESSOR IN THE DEPARTMENT OF

588
00:23:24,174 --> 00:23:28,578
PHARMAICOLOGY AND CHEMICAL

589
00:23:28,578 --> 00:23:29,012
BIOLOGY.

590
00:23:29,012 --> 00:23:30,280
SO I WILL KEEP MY INTRODUCTION

591
00:23:30,280 --> 00:23:34,618
SHORT SO YOU CAN HEAR MORE FROM

592
00:23:34,618 --> 00:23:35,185
ELISE.

593
00:23:35,185 --> 00:23:36,053
TAKE IT AWAY.

594
00:23:36,053 --> 00:23:36,887
>> THANK YOU PATTY.

595
00:23:36,887 --> 00:23:37,454
THANK YOU EVERYONE.

596
00:23:37,454 --> 00:23:38,555
THANK YOU FOR HAVING ME TODAY.

597
00:23:38,555 --> 00:23:40,090
SO I WILL TALK TODAY ABOUT A

598
00:23:40,090 --> 00:23:44,695
PROJECT WE HAVE PUBLISHED THIS

599
00:23:44,695 --> 00:23:47,030
YEAR QUITE RECENTLY THAT AIM TO

600
00:23:47,030 --> 00:23:53,770
DECIDE FURTHER DYNAM

601
00:23:53,770 --> 00:24:04,314
NAMES--NAMESSICS OF PARTNERSHIP

602
00:24:05,649 --> 00:24:05,782
PARP1

603
00:24:05,782 --> 00:24:10,620
BINDING R-LOOPS.

604
00:24:10,620 --> 00:24:12,155
--AND THEY HAVE--THEY ARE

605
00:24:12,155 --> 00:24:13,857
FAVORED BY NEGATIVE

606
00:24:13,857 --> 00:24:15,926
[INDISCERNIBLE] AND ALSO GCQs

607
00:24:15,926 --> 00:24:17,227
AND THEY ARE PHYSIOLOGY CALLED

608
00:24:17,227 --> 00:24:20,330
ROLLS IN THE CELLS, BUT

609
00:24:20,330 --> 00:24:24,067
UNFORTUNATELY THEY CAN BE

610
00:24:24,067 --> 00:24:25,001
SIGNIFICANT FOR GENOMIC

611
00:24:25,001 --> 00:24:25,302
INSTABILITY.

612
00:24:25,302 --> 00:24:26,069
THEREFORE THERE ARE MANY

613
00:24:26,069 --> 00:24:27,604
MECHANISMS THAT THE CELLS

614
00:24:27,604 --> 00:24:29,573
EVOLVED TO EITHER PREVENT THE

615
00:24:29,573 --> 00:24:31,041
FORMATION OR RESOLVING THEM

616
00:24:31,041 --> 00:24:36,847
EMPLOY AND SO WHEN WE TOOK UPON

617
00:24:36,847 --> 00:24:39,382
THIS PROGEC IN 2020, WE ASKED

618
00:24:39,382 --> 00:24:40,817
THE REQUESTY, COULD PARP1 BE 1

619
00:24:40,817 --> 00:24:43,353
OF THESE FACTORS AND WE HAD SOME

620
00:24:43,353 --> 00:24:45,122
PREMISEIS FOR THAT, IN THE

621
00:24:45,122 --> 00:24:46,990
RETURN, IT HAD BEEN ASSOCIATED

622
00:24:46,990 --> 00:24:49,359
TO THE CANNED WHAT INTERACTOME

623
00:24:49,359 --> 00:24:51,394
AND BEEN FOUND TO IRPT ACT WITH

624
00:24:51,394 --> 00:24:53,497
THE DNA, RNA LEGALLIC CASE, AND

625
00:24:53,497 --> 00:24:56,266
THEN LATER ON WHEN WE WERE ON

626
00:24:56,266 --> 00:24:58,268
UNDERTAKING THIS PROJECT,

627
00:24:58,268 --> 00:24:59,469
ANOTHER EVIDENT CAME FROM

628
00:24:59,469 --> 00:25:03,507
LITERATURE SHOWING THAT PARP1

629
00:25:03,507 --> 00:25:07,344
INTERACTS AND COAALATE WITH BP1

630
00:25:07,344 --> 00:25:07,978
AND [INDISCERNIBLE] TREATMENT

631
00:25:07,978 --> 00:25:10,580
AND DURING THE VISION OF HER

632
00:25:10,580 --> 00:25:12,082
PAPER IN [INDISCERNIBLE] WE WERE

633
00:25:12,082 --> 00:25:16,419
VERY HAPPY TO SEE THAT ACTUALLY

634
00:25:16,419 --> 00:25:21,358
ANOTHER TEAM SHOWED THAT THE

635
00:25:21,358 --> 00:25:22,292
[INDISCERNIBLE] IS SYNTHESIZED

636
00:25:22,292 --> 00:25:23,160
BY PARP1.

637
00:25:23,160 --> 00:25:25,428
SO THIS REASSURED US THIS OUR

638
00:25:25,428 --> 00:25:27,330
HYPOTHESIS THAT PARP1 CAN DO IT

639
00:25:27,330 --> 00:25:28,498
BY OWN.

640
00:25:28,498 --> 00:25:31,034
ALSO BECAUSE PARP1 NOT ONLY

641
00:25:31,034 --> 00:25:32,569
BINDENTIOUS FICIENTLY TO DNA

642
00:25:32,569 --> 00:25:34,738
BREAKS ABOUT YOU ALSO TO

643
00:25:34,738 --> 00:25:36,907
SECONDARY STRUCTURES AND IT SHOW

644
00:25:36,907 --> 00:25:38,308
INTEREST THIS KNOWING MORE ABOUT

645
00:25:38,308 --> 00:25:43,413
THAT, WE HAVE  PUBLISHED A REVIW

646
00:25:43,413 --> 00:25:46,049
IN JMB SO I INVITE TO YOU READ

647
00:25:46,049 --> 00:25:47,651
MORE ABOUT THAT.

648
00:25:47,651 --> 00:25:50,220
SO PARP1 BINDING DURING THOSE

649
00:25:50,220 --> 00:25:51,054
STRUCTURES DIRECTLY ACTIVATE

650
00:25:51,054 --> 00:25:51,588
PARP1.

651
00:25:51,588 --> 00:25:53,824
SO THE QUESTION WAS COULD PARP1

652
00:25:53,824 --> 00:25:58,061
DIRECTLY RECOGNIZE THE STRUCTURE

653
00:25:58,061 --> 00:25:59,763
AND BE ACTIVITIED TO THE BINDING

654
00:25:59,763 --> 00:26:00,063
STRUCTURES.

655
00:26:00,063 --> 00:26:01,631
SO THIS IS A PAPER WE JUST

656
00:26:01,631 --> 00:26:06,603
PUBLISHED IN YEN --JANUARY OF

657
00:26:06,603 --> 00:26:10,040
THIS YEAR.

658
00:26:10,040 --> 00:26:11,675
WE COMBINED BIOLOGY AND

659
00:26:11,675 --> 00:26:12,976
MICROSCOPY AND I WILL GO OVER

660
00:26:12,976 --> 00:26:13,443
THE DATA.

661
00:26:13,443 --> 00:26:17,280
SO 1 THING THAT WE DID IS THAT

662
00:26:17,280 --> 00:26:19,282
WE USE A PLAZ MIDS MIDTHAT WAS

663
00:26:19,282 --> 00:26:21,685
GENERATED IN THE LAB AND KINDLY

664
00:26:21,685 --> 00:26:23,820
PROVIED TO US BY [INDISCERNIBLE]

665
00:26:23,820 --> 00:26:29,059
WHICH IS OF COURSE THE PFCAMAN.

666
00:26:29,059 --> 00:26:30,927
IT'S A MOUSE GENE THAT IS PRONE

667
00:26:30,927 --> 00:26:37,000
TO  [INDISCERNIBLE] MUTATION, IT

668
00:26:37,000 --> 00:26:38,969
WILL GO THROUGH INVITRO

669
00:26:38,969 --> 00:26:41,371
TRANSCRIPTION, IT FORMS AN--IT

670
00:26:41,371 --> 00:26:47,077
FORMS RNA AND IT FORMS THIS--AS

671
00:26:47,077 --> 00:26:49,246
WE CAN VERIFY BY GEL HERE WE CAN

672
00:26:49,246 --> 00:26:51,848
SEE AFTER THE TRANSCRIPTION,

673
00:26:51,848 --> 00:26:53,183
THIS SHIFT IN THE MIGRATION,

674
00:26:53,183 --> 00:26:55,652
HERE YOU CAN SEE THAT IF WE

675
00:26:55,652 --> 00:26:57,320
DIGEST WITH RNA, WE CAN GET RID

676
00:26:57,320 --> 00:26:59,055
OF THE FREE RNA BUT NOT THE

677
00:26:59,055 --> 00:27:04,494
SHIFT, BUT THEN IF WE DIGEST,

678
00:27:04,494 --> 00:27:06,496
WHICH DIGEST THE RNA IN THE

679
00:27:06,496 --> 00:27:12,202
HYBRID OF THE RNA- DNA.

680
00:27:12,202 --> 00:27:13,837
THE SHIFT IS ABOLERBED HERE AND

681
00:27:13,837 --> 00:27:15,605
GOES BACK TO REGULAR PLAZ MIDS

682
00:27:15,605 --> 00:27:15,939
MIDMIGRATION.

683
00:27:15,939 --> 00:27:18,742
SO WE USE THIS SPECIMEN TO FIRST

684
00:27:18,742 --> 00:27:20,577
INTERROGATE THE BINDING OF PARP1

685
00:27:20,577 --> 00:27:24,147
BY FM AND HERE WE TEAMED UP WITH

686
00:27:24,147 --> 00:27:27,851
THE COLLABORATOR [INDISCERNIBLE]

687
00:27:27,851 --> 00:27:31,021
AT NCSU WHO HAVE THIS AFM, SO

688
00:27:31,021 --> 00:27:31,988
THEY USE THIS PLAZ MIDS MIDTHAT

689
00:27:31,988 --> 00:27:36,326
WE HAVE BUT HERE WE DIGEST THE

690
00:27:36,326 --> 00:27:37,928
PLAZ MIDS MIDWITH APL1 THAT

691
00:27:37,928 --> 00:27:39,763
GENERATES FRAGMENT IN HAD WE

692
00:27:39,763 --> 00:27:41,765
KNOW THAT THERE ARE FORMS AROUND

693
00:27:41,765 --> 00:27:44,768
40% FORM THE CLOSEST DMA END

694
00:27:44,768 --> 00:27:51,441
WHICH WE CAN VERIFY HERE, BY

695
00:27:51,441 --> 00:27:54,077
AFM, AND YOU CAN SEE HERE THAT

696
00:27:54,077 --> 00:27:56,680
BY COLLECTING AFM HEIGHTS THAT

697
00:27:56,680 --> 00:27:59,582
THIS LOOP IS AROUND 40% FROM THE

698
00:27:59,582 --> 00:28:00,517
CLOSEST DNA.

699
00:28:00,517 --> 00:28:05,155
WE THEN ENCUE BAITED PARP1 WITH

700
00:28:05,155 --> 00:28:06,790
THE SAME FRAGMENT BUT WITHOUT

701
00:28:06,790 --> 00:28:10,961
TRANSCRIPTION AND AS EXPECTED

702
00:28:10,961 --> 00:28:13,096
PARP1 CAN BIND AT DNA ENDS,

703
00:28:13,096 --> 00:28:15,999
WHICH IS QUITE HIGH BUT ALSO

704
00:28:15,999 --> 00:28:18,435
HERE ON THE FRAGMENT HERE BUT

705
00:28:18,435 --> 00:28:25,709
WHEN WE INCUBATE PARP1 WITH THE

706
00:28:25,709 --> 00:28:26,876
LOOP AFTER RVT, CAN YOU SEE

707
00:28:26,876 --> 00:28:30,380
THERE IS A SHIFT OF PARTNERSHIP

708
00:28:30,380 --> 00:28:33,116
BINDING AROUND 40% OF PROCESS

709
00:28:33,116 --> 00:28:35,752
UPON BINDING TO THE EXTREMITIES

710
00:28:35,752 --> 00:28:37,687
WHICH SHOWS THAT PARP1 INDEED

711
00:28:37,687 --> 00:28:40,457
BINDS TO ALL STRUCTURES HERE.

712
00:28:40,457 --> 00:28:44,094
SO, WE ALSO WANTED TO CHECK IN

713
00:28:44,094 --> 00:28:47,263
CELLS IF PRARP 1 COULD BIND IN

714
00:28:47,263 --> 00:28:50,600
OUR LOOP FORMING SIDES, SO FOR

715
00:28:50,600 --> 00:28:53,670
THAT WE ESTABLISH A CELL LINE IN

716
00:28:53,670 --> 00:28:55,605
WHICH WE CAN OVEREXPRESS USING

717
00:28:55,605 --> 00:28:58,008
DOCKS O PSYCHE LYNN, EITHER THE

718
00:28:58,008 --> 00:29:00,343
WILD-TYPE OR THE MUTANT OVER

719
00:29:00,343 --> 00:29:03,246
EXPRESSING THE MUTANT LEADS TO

720
00:29:03,246 --> 00:29:04,014
INCREASE OF R-LOOP.

721
00:29:04,014 --> 00:29:06,716
SO WE USE OUR CELL LINE HERE TO

722
00:29:06,716 --> 00:29:08,885
INTERROGATE THE BINDING OF PARP1

723
00:29:08,885 --> 00:29:11,488
ON THE SITES BY THE LIGATION

724
00:29:11,488 --> 00:29:12,489
ASSAY USING THE [INDISCERNIBLE]

725
00:29:12,489 --> 00:29:15,458
WHICH IS THE DNA, RNA HYBRID,

726
00:29:15,458 --> 00:29:17,727
AND USING PARP1 ANTIBODY, HERE

727
00:29:17,727 --> 00:29:23,933
ARE THE ANTIBODY CONTROLS, FOR

728
00:29:23,933 --> 00:29:27,337
THE EXPERIMENT, WE NEED TO

729
00:29:27,337 --> 00:29:28,738
PRETREAT OURSELVES WITH RNA-1

730
00:29:28,738 --> 00:29:31,408
AND 3, WHICH GETS RID OF THE

731
00:29:31,408 --> 00:29:33,810
SINGLE STRANDED ASK DOUBLE

732
00:29:33,810 --> 00:29:35,211
STRANDED RNA WHICH HAS SOME

733
00:29:35,211 --> 00:29:37,781
AFFINITY SO THAT MAKES OUR

734
00:29:37,781 --> 00:29:38,314
EXPERIMENT MORE SPECIFIC.

735
00:29:38,314 --> 00:29:43,620
AND YOU CAN SEE HERE THAT IF WE

736
00:29:43,620 --> 00:29:49,292
USE THE CELL LINE EXPRESSING THE

737
00:29:49,292 --> 00:29:50,460
WILD-TYPE, WE HOWEVER, YOU CAN

738
00:29:50,460 --> 00:29:52,362
SEE HERE THAT'S QUANTIFIED HERE,

739
00:29:52,362 --> 00:29:55,098
AN INCREASE OF FOCI FORM BETWEEN

740
00:29:55,098 --> 00:29:57,634
PARP1 AND INDEX EABT BODY, WHICH

741
00:29:57,634 --> 00:30:02,205
SHOWS THAT PARP1 IS ASSOCIATED

742
00:30:02,205 --> 00:30:03,139
TO THE ALLUMFORMING SITES.

743
00:30:03,139 --> 00:30:05,909
AND WE VALIDATED THIS IN A CELL

744
00:30:05,909 --> 00:30:08,344
THAT WAS DEPLETED IN RNA

745
00:30:08,344 --> 00:30:11,681
[INDISCERNIBLE] AND ALSO ANOTHER

746
00:30:11,681 --> 00:30:12,348
FACTOR, RESOLUTION FACTOR.

747
00:30:12,348 --> 00:30:15,518
SO OF COURSE, OUR NEXT QUESTION

748
00:30:15,518 --> 00:30:17,921
WAS IS PARP1 ACTIVITY TRIGGERED

749
00:30:17,921 --> 00:30:18,254
UPON BINDING.

750
00:30:18,254 --> 00:30:21,958
SO TO DO THAT, WE DID THE PARP1

751
00:30:21,958 --> 00:30:23,793
OR 2 MODIFICATION ASSAY INVITRO,

752
00:30:23,793 --> 00:30:26,162
SO FOR THAT WE INCUBATE PARP1

753
00:30:26,162 --> 00:30:30,100
WITH THE CO FACTOR NAD, WITH DMA

754
00:30:30,100 --> 00:30:31,701
THAT HAS SINGLE STRAND BREAKS

755
00:30:31,701 --> 00:30:32,802
WHICH IS POSITIVE CONTROL WITH

756
00:30:32,802 --> 00:30:35,338
THE PLAZ MIDS MID, WITH IT TIME

757
00:30:35,338 --> 00:30:37,407
REMAINS CIRCULAR TO AVOID THE

758
00:30:37,407 --> 00:30:39,709
PRESENCE OF DNA THAT WOULD

759
00:30:39,709 --> 00:30:41,578
ACTIVATE FOR PARP1 AND THEN WITH

760
00:30:41,578 --> 00:30:43,079
THE PLAZ MIDS MIDTHAT HAVE THE

761
00:30:43,079 --> 00:30:47,283
LOOP, AND YOU CAN SEE HERE THAT

762
00:30:47,283 --> 00:30:50,954
UPON INCUBATION OF PARP1 AND THE

763
00:30:50,954 --> 00:30:53,289
SUBSTRATE THAT WE HAVE HIGH

764
00:30:53,289 --> 00:30:54,424
SYNTHESIS OF [INDISCERNIBLE] IN

765
00:30:54,424 --> 00:30:56,092
POSITIVE CONTROL, A LITTLE BIT

766
00:30:56,092 --> 00:31:02,265
IN THE  PLASMID THAT DOESN'T HAE

767
00:31:02,265 --> 00:31:04,100
THE LOOP, WHICH MAY HAVE SOME

768
00:31:04,100 --> 00:31:05,768
STRUCTURES OR SOME NICKS, BUT

769
00:31:05,768 --> 00:31:08,972
IMPORTANTLY WE SEE A SIGNIFICANT

770
00:31:08,972 --> 00:31:10,807
INCREASE OF [INDISCERNIBLE] WHEN

771
00:31:10,807 --> 00:31:13,443
WE INCUBATE WITH THE PLASMID

772
00:31:13,443 --> 00:31:15,178
THAT CARRIES THE R-LOOP.

773
00:31:15,178 --> 00:31:19,949
SO WE ALSO VALIDATED THAT BY IF

774
00:31:19,949 --> 00:31:21,251
IN OUR RETURNED CELL LINE WHERE

775
00:31:21,251 --> 00:31:28,224
WE SHOW AN INCREASE OF

776
00:31:28,224 --> 00:31:28,725
[INDISCERNIBLE].

777
00:31:28,725 --> 00:31:30,994
ALSO WE WERE CURIOUS TO SEE HOW

778
00:31:30,994 --> 00:31:34,097
PARP1 LOOKS BY CURATED FM, SO

779
00:31:34,097 --> 00:31:35,598
HERE WE HAVE TO CHANGE OUR

780
00:31:35,598 --> 00:31:37,534
PARAMETERS TO SLOW DOWN THE

781
00:31:37,534 --> 00:31:38,201
REACTION, AGAIN, TALK LATER

782
00:31:38,201 --> 00:31:41,938
ABOUT THE REASON WHY, BUT

783
00:31:41,938 --> 00:31:43,873
IMPORTANTLY WE SOLD THAT WE HAVE

784
00:31:43,873 --> 00:31:47,210
PATHWAY 1, WE HAVE BRANCH, THAT

785
00:31:47,210 --> 00:31:49,078
HAS--THAT IS HOW ABOUT A 40%

786
00:31:49,078 --> 00:31:50,947
FROM THE CLOSEST DNA, WHICH

787
00:31:50,947 --> 00:31:56,352
SHOWS IN THE PARAALATED TOP 1 IN

788
00:31:56,352 --> 00:31:57,120
THE R-LOOPS.

789
00:31:57,120 --> 00:31:59,155
SO HERE SO FAR WE FIND THIS

790
00:31:59,155 --> 00:32:00,356
BINDING IN THE LOOP AND THE

791
00:32:00,356 --> 00:32:01,524
ACTIVITY, BUT WE WANT TO

792
00:32:01,524 --> 00:32:04,961
UNDERSTAND THE IMPACT OF PARP1

793
00:32:04,961 --> 00:32:07,597
DELETION AND INHIBITION ON

794
00:32:07,597 --> 00:32:08,097
[INDISCERNIBLE] GENOME

795
00:32:08,097 --> 00:32:08,364
STABILITY.

796
00:32:08,364 --> 00:32:13,369
ONE THING THAT WE DID, IS DNA

797
00:32:13,369 --> 00:32:14,737
HYBRID BLAD BLOT IN OUR CELL

798
00:32:14,737 --> 00:32:17,373
LINE THAT WE KNOCKED OUT IN

799
00:32:17,373 --> 00:32:23,213
WHICH WE KNOCKED OUT PARP1 BUT

800
00:32:23,213 --> 00:32:24,881
WE ALSO REEXPRESSED PARP1 TO SEE

801
00:32:24,881 --> 00:32:26,316
IF IT RESCUES THE PHENOTYPE.

802
00:32:26,316 --> 00:32:27,717
YOU CAN SEE THE ACCUMULATION AND

803
00:32:27,717 --> 00:32:30,386
YOU CAN SEE THE BLOOD USING THE

804
00:32:30,386 --> 00:32:31,921
ANTIBODY WITH A SIGNIFICANT

805
00:32:31,921 --> 00:32:33,890
INCREASE OF RLOOP LEVEL HERE IN

806
00:32:33,890 --> 00:32:38,861
THE PARP1 CELLS WHICH IS RESCUED

807
00:32:38,861 --> 00:32:40,630
WHEN WE OVEREXPRESS THE PARP1

808
00:32:40,630 --> 00:32:42,932
WILD-TYPE WHICH SHOWS THAT IF WE

809
00:32:42,932 --> 00:32:45,435
REMOVE PARP1, THEN THERE IS

810
00:32:45,435 --> 00:32:47,904
ACCUMULATION OF DNA HYBRID, DMA

811
00:32:47,904 --> 00:32:50,206
RNA HYBRID IN CELLS.

812
00:32:50,206 --> 00:32:52,609
OF COURSE WE ALSO REPEATED THAT

813
00:32:52,609 --> 00:32:54,644
WE STOP 1 SiRNA AS WELL.

814
00:32:54,644 --> 00:32:56,246
FINALLY, WHAT'S THE IMPACT OF

815
00:32:56,246 --> 00:32:59,215
IPT GREATER HIB THORSITION, TOP

816
00:32:59,215 --> 00:33:02,619
1 INHIBITION, ALSO LED TO AN

817
00:33:02,619 --> 00:33:04,554
INCREASE OF X96 STAINING IN OUR

818
00:33:04,554 --> 00:33:08,992
CELL LINE THAT ALSO THAT

819
00:33:08,992 --> 00:33:10,827
EXPRESSES THE WILD-TYPE RNAH1.

820
00:33:10,827 --> 00:33:12,662
YOU CAN SEE THE INCREASE.

821
00:33:12,662 --> 00:33:14,230
THAT'S QUANTIFIED HERE AND

822
00:33:14,230 --> 00:33:15,632
THAT'S ABOLERBED WHEN WE

823
00:33:15,632 --> 00:33:20,303
PRETREAT THE CELLS WITH RNSH, TO

824
00:33:20,303 --> 00:33:22,705
FIX THE CELLS IT SHOW THIS IS

825
00:33:22,705 --> 00:33:24,374
SPECIFIC FORM DURING THE HYBRID

826
00:33:24,374 --> 00:33:26,342
ACCUMULATION AND FINALLY WE LOOK

827
00:33:26,342 --> 00:33:28,211
THEA GENOMIC INSTABILITY UPON

828
00:33:28,211 --> 00:33:29,712
PARP1 INHIBITION BY FOLLOWING

829
00:33:29,712 --> 00:33:35,685
THE FORMATION, WE DID THAT IN

830
00:33:35,685 --> 00:33:38,154
BOTH CELLS, IN S-PHASE CELLS AND

831
00:33:38,154 --> 00:33:39,622
NONS-PHASE CELLS TO SEE IF WE

832
00:33:39,622 --> 00:33:41,491
HAD IPT GREATER STABILITY ONLY

833
00:33:41,491 --> 00:33:42,992
LINKED TO REPLICATION STRESS.

834
00:33:42,992 --> 00:33:46,729
AND WHAT WE SAW IS THAT PARP1

835
00:33:46,729 --> 00:33:47,964
INHIBITION OF COURSE AS EXPECTED

836
00:33:47,964 --> 00:33:50,967
LEADS TO AN INCREASE OF GAMMA H2

837
00:33:50,967 --> 00:33:54,337
X IN BOTH CELL TYPES.

838
00:33:54,337 --> 00:33:58,041
AND WE SAW THAT ACCUMULATION OF

839
00:33:58,041 --> 00:34:00,276
R-LOOPS USING DOCKSY PSYCHE LINE

840
00:34:00,276 --> 00:34:04,247
ALSO LEADING TO ACCUMULATION OF

841
00:34:04,247 --> 00:34:06,149
R-LOOPS HERE BUT ONLY IN S-PHASE

842
00:34:06,149 --> 00:34:07,850
CELLS WHICH IS EXPECTED BECAUSE

843
00:34:07,850 --> 00:34:09,252
OF THE COLLISION BETWEEN THE

844
00:34:09,252 --> 00:34:11,454
R-LOOP AND THE REPLICATION

845
00:34:11,454 --> 00:34:11,754
MACHINERY.

846
00:34:11,754 --> 00:34:13,423
WHAT WAS INTERESTING IS THAT WE

847
00:34:13,423 --> 00:34:16,092
SHOW WHEN WE COMBINE DON,

848
00:34:16,092 --> 00:34:17,527
XYPSYCHE LINE AND INHIBITOR WE

849
00:34:17,527 --> 00:34:20,496
OBSERVE AN INCREASE IN BOTH

850
00:34:20,496 --> 00:34:23,366
S-PHASE AND NONS-PHASE CELLS,

851
00:34:23,366 --> 00:34:27,203
WHICH SHOWS ACTUALLY IT IS NOT

852
00:34:27,203 --> 00:34:28,504
ONLY CHARACTERIZED IN S-PHASE

853
00:34:28,504 --> 00:34:31,207
CELLS WHEN YOU INHIBIT PATHWAY.

854
00:34:31,207 --> 00:34:34,077
SO PARP1 HAS ENDED THE ROLE IN

855
00:34:34,077 --> 00:34:37,580
OUR [INDISCERNIBLE] IN

856
00:34:37,580 --> 00:34:38,514
PRESERVING GENOMIC INSTABILITY

857
00:34:38,514 --> 00:34:41,851
OF A DNA HYBRID AND WE OBTAINED

858
00:34:41,851 --> 00:34:45,288
THAT SIMILARLY IN HELIX CELLS,

859
00:34:45,288 --> 00:34:46,956
SO THIS IS A QUICK SUMMARY OF

860
00:34:46,956 --> 00:34:48,558
THE PAPER THAT'S BEEN PUBLISHED.

861
00:34:48,558 --> 00:34:50,259
I INVITE YOU TO READ OVER FOR

862
00:34:50,259 --> 00:34:51,260
MORE DETAILS BUT OF COURSE NOW

863
00:34:51,260 --> 00:34:52,662
WE HAVE, I WANTED TO TALK A

864
00:34:52,662 --> 00:34:56,165
LITTLE BIT ABOUT OUR NEXT STEPS.

865
00:34:56,165 --> 00:34:57,500
AND WE HAVE MAYBE 3 OF THOSE

866
00:34:57,500 --> 00:34:58,835
QUESTIONS, SO WHAT ARE THE

867
00:34:58,835 --> 00:35:01,204
DOMAINS OF PARP1 THAT ARE

868
00:35:01,204 --> 00:35:04,407
INVOLVED IN THE BINDING, WHERE

869
00:35:04,407 --> 00:35:05,708
IT BINDING BECAUSE IT'S RATHER

870
00:35:05,708 --> 00:35:07,744
LARGE STRUCTURE AND TO DO WHAT?

871
00:35:07,744 --> 00:35:09,679
AND SO, WE ARE NOW RACING THIS

872
00:35:09,679 --> 00:35:11,614
FIRST QUESTION ABOUT THE DOMAINS

873
00:35:11,614 --> 00:35:15,151
INVOLVED, SO THERE ARE A LOT OF

874
00:35:15,151 --> 00:35:17,286
WORK, IN THE P A RPFIELD THAT

875
00:35:17,286 --> 00:35:20,256
SHOWS THAT PARP1 BINDING ON THE

876
00:35:20,256 --> 00:35:21,824
CANNONICLE SUBSTRATE ON THE

877
00:35:21,824 --> 00:35:23,926
SINGLE AND DOUBLE STRAND BREAKS

878
00:35:23,926 --> 00:35:25,628
THAT DEMON TRAIT THAT THOSE

879
00:35:25,628 --> 00:35:27,063
DOMAINS, THE AND THE DOUBLE GR

880
00:35:27,063 --> 00:35:28,731
BYPASSED ON THE BREAK AND THE

881
00:35:28,731 --> 00:35:31,801
REMODELING OF THESE DOMAIN WILL

882
00:35:31,801 --> 00:35:35,138
ALLOW FOR THE REMODELING OF THE

883
00:35:35,138 --> 00:35:36,739
CATALYTIC DOMAIN AND PARP1

884
00:35:36,739 --> 00:35:37,540
ACTIVATION.

885
00:35:37,540 --> 00:35:39,175
SO SINCE WE SEE PARP1 ACTIVATION

886
00:35:39,175 --> 00:35:40,843
ON OUR LOOP AND STRUCTURE, WE

887
00:35:40,843 --> 00:35:43,179
WONDERED IF SOME OF THOSE

888
00:35:43,179 --> 00:35:44,213
DOMAINS ARE ALSO INVOLVED, BUT

889
00:35:44,213 --> 00:35:47,784
FIRST TO DO THAT, WE

890
00:35:47,784 --> 00:35:49,552
COLLABORATED, WE ARE

891
00:35:49,552 --> 00:35:50,420
COLLABORATING WITH BEN

892
00:35:50,420 --> 00:35:52,722
[INDISCERNIBLE] LAB HERE AT

893
00:35:52,722 --> 00:35:52,955
HILLMAN.

894
00:35:52,955 --> 00:35:56,726
AND WITH THE HELP OF MATT SHAY

895
00:35:56,726 --> 00:35:58,127
AND [INDISCERNIBLE] IN HIS LAB,

896
00:35:58,127 --> 00:36:03,966
WE HAVE FINALLY COME UP WITH A

897
00:36:03,966 --> 00:36:05,835
SITUATION ON THE

898
00:36:05,835 --> 00:36:06,302
[INDISCERNIBLE].

899
00:36:06,302 --> 00:36:13,109
AND SO WE ARE FOLLOWING A

900
00:36:13,109 --> 00:36:14,110
PROTOCOL.

901
00:36:14,110 --> 00:36:15,411
AND IT'S A 5 CHAMBER FLOW CELL

902
00:36:15,411 --> 00:36:17,947
SO WE CAN FLOW THE COMPLEMENT,

903
00:36:17,947 --> 00:36:20,917
SO FIRST WE FLOW THE

904
00:36:20,917 --> 00:36:21,284
[INDISCERNIBLE].

905
00:36:21,284 --> 00:36:23,553
AND THEN WE FLOW THE SUBSTRATE,

906
00:36:23,553 --> 00:36:27,156
WE FLOW BUFFER AND THEN WE FLOW

907
00:36:27,156 --> 00:36:29,158
OUR CELL EXTRACT THAT CONTAINS

908
00:36:29,158 --> 00:36:30,460
THE FLUORESCENTLY TAGGED PROTEIN

909
00:36:30,460 --> 00:36:30,893
OF INTEREST.

910
00:36:30,893 --> 00:36:33,296
AND THEN WE CAN FOLLOW THE

911
00:36:33,296 --> 00:36:35,598
BINDING ON OUR SUBSTRATE THAT

912
00:36:35,598 --> 00:36:36,866
TRANSLATE INTO THIS CHEMO GRAPH

913
00:36:36,866 --> 00:36:41,370
HERE POSITION HERE AND IN TIME.

914
00:36:41,370 --> 00:36:43,973
SO WHAT WE DID AND WHAT THEY

915
00:36:43,973 --> 00:36:48,811
DID, IS THAT SHE USED THE GENE

916
00:36:48,811 --> 00:36:52,148
THAT SHE HAD IN THE PLAZ MIDS

917
00:36:52,148 --> 00:36:54,817
MIDWHICH ALLOW FOR DIGESTION

918
00:36:54,817 --> 00:36:57,553
WITH BBS 1 AND THE BINDING OF

919
00:36:57,553 --> 00:37:00,256
THOSE [INDISCERNIBLE] HANDLE

920
00:37:00,256 --> 00:37:02,291
THAT ALLOWS R-LOOP SUBSTRATE TO

921
00:37:02,291 --> 00:37:04,827
BE BOUND BETWEEN THE BEADS OF

922
00:37:04,827 --> 00:37:08,130
THE SIT REP AND WHAT ALSO--WHAT

923
00:37:08,130 --> 00:37:12,602
THE NATALY DID HERE IS SHE DID

924
00:37:12,602 --> 00:37:13,769
INVITRO TRANSCRIPTION USING ATP

925
00:37:13,769 --> 00:37:15,638
WHICH ALLOWS US TO VISUALIZE THE

926
00:37:15,638 --> 00:37:18,608
LOOP DIRECTLY ON THE MICROSCOPE.

927
00:37:18,608 --> 00:37:23,145
SO I'M JUST GOING OVER SOME

928
00:37:23,145 --> 00:37:26,916
SMALL CAMERA GRAPH WE OBTAINED

929
00:37:26,916 --> 00:37:27,650
WITH THE WILD-TYPE ENZYME.

930
00:37:27,650 --> 00:37:29,252
Y SOPHISTICATED YOU CAN SEE

931
00:37:29,252 --> 00:37:31,187
PARP1 BYPASSEDS OR IT IS VERY

932
00:37:31,187 --> 00:37:32,822
STABLE OR CAN BE VERY MOBILE

933
00:37:32,822 --> 00:37:37,960
ALONG THE D, INA, AND WE CAME UP

934
00:37:37,960 --> 00:37:39,462
WITH CUMULATIVE [INDISCERNIBLE]

935
00:37:39,462 --> 00:37:43,533
DISTRIBUTED HERE AND THE HALF

936
00:37:43,533 --> 00:37:44,567
LIFE AROUND 5.2-SECOND WHICH IS

937
00:37:44,567 --> 00:37:46,502
VERY CLOSE TO THE LIFETIME

938
00:37:46,502 --> 00:37:49,038
OBSERVED ON THE NICK DNA.

939
00:37:49,038 --> 00:37:52,208
SO WE ARE NOW ADDRESSING THE

940
00:37:52,208 --> 00:37:55,011
IMPACT OF SOME MUTATION, SOME

941
00:37:55,011 --> 00:37:57,079
PARP1 MUTANTS, OLDER MUTANT WE

942
00:37:57,079 --> 00:37:59,181
ARE NOT TESTING SO STAY TUNED

943
00:37:59,181 --> 00:38:01,617
FOR WHAT WILL HAPPEN WITH THOSE

944
00:38:01,617 --> 00:38:03,219
MUTANT AND SO THIS IS TIME FOR

945
00:38:03,219 --> 00:38:07,256
ME TO THANK MY LAB, AND NATALY,

946
00:38:07,256 --> 00:38:10,459
MY, Ph.D. STUDENT WHO LED TO

947
00:38:10,459 --> 00:38:12,328
PROG EXPECT GRADUATED LAST WEEK

948
00:38:12,328 --> 00:38:15,264
AND OUR COLLABORATORS OF COURSE,

949
00:38:15,264 --> 00:38:17,567
THE LAB FOR THE [INDISCERNIBLE]

950
00:38:17,567 --> 00:38:19,502
WE ARE LEADING RIGHT NOW.

951
00:38:19,502 --> 00:38:20,736
AND OF COURSE, THE FUNDING AND

952
00:38:20,736 --> 00:38:30,980
THANK YOU FOR YOUR ATTENTION.

953
00:38:30,980 --> 00:38:31,914
>> THANK YOU.

954
00:38:31,914 --> 00:38:33,783
YOUR FIRST QUESTION IS ROB ASKS

955
00:38:33,783 --> 00:38:37,119
IF THERE IS A FORMATION FOR

956
00:38:37,119 --> 00:38:38,621
PROCESSING AND ROLE PROCESSING?

957
00:38:38,621 --> 00:38:41,490
AND THE SECOND QUESTION IS ABOUT

958
00:38:41,490 --> 00:38:42,124
PARTNERSHIP 2?

959
00:38:42,124 --> 00:38:44,560
>> SO ABOUT P A RP2, IT
WOULD BE

960
00:38:44,560 --> 00:38:46,395
INTERESTING WHEN WE KNOW WHAT

961
00:38:46,395 --> 00:38:49,432
DOMAIN ARE INVOLVED WE MAY BE

962
00:38:49,432 --> 00:38:51,801
ABLE TO ALSO TEST P A RP2, BUT

963
00:38:51,801 --> 00:38:53,703
ALSO SEE THAT P A RP2 IS

964
00:38:53,703 --> 00:38:55,438
INVOLVED THERE BECAUSE AS YOU

965
00:38:55,438 --> 00:38:57,173
KNOW THE MODEL IS BIEBDING TO

966
00:38:57,173 --> 00:38:57,974
THE [INDISCERNIBLE] IS VERY

967
00:38:57,974 --> 00:39:00,242
DIFFERENT BECAUSE IT DOESN'T

968
00:39:00,242 --> 00:39:02,011
HAVE THE SAME DNA BINDING

969
00:39:02,011 --> 00:39:03,012
DOMAIN, BUT THIS IS SOMETHING WE

970
00:39:03,012 --> 00:39:05,081
WILL LIKE TO TEST FOR SURE.

971
00:39:05,081 --> 00:39:05,982
IT SEEMS INTERESTING.

972
00:39:05,982 --> 00:39:08,284
AND THEN THE SECOND QUESTION, IS

973
00:39:08,284 --> 00:39:12,154
THE [INDISCERNIBLE]OT END OF

974
00:39:12,154 --> 00:39:12,455
C-TERMINUS?

975
00:39:12,455 --> 00:39:14,724
I THINK IT'S ON THE END TERMINUS

976
00:39:14,724 --> 00:39:18,394
IN I REMEMBER, YEAH.

977
00:39:18,394 --> 00:39:19,495
>> SO I HAVE A QUICK
QUESTION

978
00:39:19,495 --> 00:39:20,863
EMPLOY I DON'T SEE OTHERS IN THE

979
00:39:20,863 --> 00:39:22,598
CHAT YET.

980
00:39:22,598 --> 00:39:24,467
DO YOU THINK THAT--SO I LIKED

981
00:39:24,467 --> 00:39:26,035
YOUR QUESTION ABOUT HOW IS PARP1

982
00:39:26,035 --> 00:39:27,703
DOING THIS, DO YOU THINK THAT

983
00:39:27,703 --> 00:39:30,006
THE PROCESS OF THIS MIGHT BE

984
00:39:30,006 --> 00:39:31,574
ENOUGH TO DISLODGE THE R-LOOP OR

985
00:39:31,574 --> 00:39:35,511
DO YOU THINK THAT PARTNERSHIP IS

986
00:39:35,511 --> 00:39:37,613
ACTUALLY RECRUITING SPECIFIC

987
00:39:37,613 --> 00:39:39,815
HELIC CASE OR SEVERAL FOR THIS

988
00:39:39,815 --> 00:39:39,982
IN.

989
00:39:39,982 --> 00:39:42,818
>> THE HIGH THROUGH PUT IS
THE

990
00:39:42,818 --> 00:39:45,554
FIRST ACTIVITY OF PARP1 ALLOWED

991
00:39:45,554 --> 00:39:47,757
FACTORS THAT ARE LESS ABUNDANT

992
00:39:47,757 --> 00:39:50,726
COMPARED TO PARP1 IN THE CELL TO

993
00:39:50,726 --> 00:39:53,496
QUICKLY GET THE R-LOOP

994
00:39:53,496 --> 00:39:53,763
BASICALLY.

995
00:39:53,763 --> 00:39:54,897
LIKE PARP1 WOULD DO ON THE

996
00:39:54,897 --> 00:39:57,767
BREAK, SO HAD IS WHAT WE'RE

997
00:39:57,767 --> 00:39:59,068
INTERROGATING WITH OUR

998
00:39:59,068 --> 00:39:59,869
[INDISCERNIBLE] THAT HOPEFULLY

999
00:39:59,869 --> 00:40:01,537
WILL BE FUNDED SOON.

1000
00:40:01,537 --> 00:40:02,104
[LAUGHTER]

1001
00:40:02,104 --> 00:40:02,772
>> DPRAIT.

1002
00:40:02,772 --> 00:40:03,906
AND 1 QUESTION FROM BRETT

1003
00:40:03,906 --> 00:40:04,640
[INDISCERNIBLE] AND THIS WILL

1004
00:40:04,640 --> 00:40:06,308
HAVE TO BE THE LAST 1, SO WE

1005
00:40:06,308 --> 00:40:08,778
HAVE TIME FOR THE THIRD TALK,

1006
00:40:08,778 --> 00:40:10,579
GREAT TALK, DID YOUR SUBSTRATES

1007
00:40:10,579 --> 00:40:12,548
FOR THE C-TRAP HAVE THE RNA

1008
00:40:12,548 --> 00:40:14,583
COMPONENT FOR THE R-LOOP, I MAY

1009
00:40:14,583 --> 00:40:15,384
ARE MISSED THIS.

1010
00:40:15,384 --> 00:40:18,054
>> YEAH, SO WE DO IVT
BEFORE TO

1011
00:40:18,054 --> 00:40:25,261
TETHER THE SUBSTRATE ON TO THE

1012
00:40:25,261 --> 00:40:26,896
[INDISCERNIBLE], SO THE TAIL OF

1013
00:40:26,896 --> 00:40:29,565
THE RNA IS DIGESTED WE JUST HAVE

1014
00:40:29,565 --> 00:40:32,368
THE DNA HIBRIDE AND THE

1015
00:40:32,368 --> 00:40:33,903
DISPLACED SINGLE STRANDED DNA.

1016
00:40:33,903 --> 00:40:34,303
>> GREAT.

1017
00:40:34,303 --> 00:40:34,870
RIGHT ON TIME.

1018
00:40:34,870 --> 00:40:45,314
THANK YOU SO MUCH ELISE. 

1019
00:40:47,717 --> 00:40:50,352
NTHANK YOU EMPLOY.

1020
00:40:50,352 --> 00:40:54,290
>> OKAY, GREAT DO I JUST
JUMP?

1021
00:40:54,290 --> 00:40:54,724
S.

1022
00:40:54,724 --> 00:40:59,228
>> SO NOW WE HAVE CLAUDIA
WITH

1023
00:40:59,228 --> 00:41:01,664
PENN STATE DEPARTMENT OF

1024
00:41:01,664 --> 00:41:03,332
MOLECULAR BIOLOGY.

1025
00:41:03,332 --> 00:41:04,734
SHE FINISHED HER Ph.D. IN THE

1026
00:41:04,734 --> 00:41:06,535
INSTITUTE OF BI ONY CHEMISTRY

1027
00:41:06,535 --> 00:41:08,370
AND WEPT ON TO POST DOCTORAL

1028
00:41:08,370 --> 00:41:09,672
POSITIONS AT HARVARD SCHOOL OF

1029
00:41:09,672 --> 00:41:11,640
DEPTAL MEDICINE AND THEN PENN

1030
00:41:11,640 --> 00:41:12,775
STATE WAS PROMOTED THROUGH THE

1031
00:41:12,775 --> 00:41:14,210
RAIRCHGS AT PENN STATE TO

1032
00:41:14,210 --> 00:41:15,644
RESEARCH ASSOCIATE AND ASSISTANT

1033
00:41:15,644 --> 00:41:16,579
PROFESSOR RESEARCH TRACK AND NOW

1034
00:41:16,579 --> 00:41:19,115
OF COURSE ON THE TENURED TRACK

1035
00:41:19,115 --> 00:41:22,118
AND HER WORK IS CURRENTLY FUNDED

1036
00:41:22,118 --> 00:41:24,920
BY AN NCI RO-1 FOCUSED ON PARP10

1037
00:41:24,920 --> 00:41:26,188
WITH REGARD TO REPLICATION

1038
00:41:26,188 --> 00:41:27,623
STRESS AND THAT'S GOING TO BE

1039
00:41:27,623 --> 00:41:29,592
THE FOCUS OF OUR TALK TODAY.

1040
00:41:29,592 --> 00:41:34,463
REALLY EXCITED TO HEAR YOUR WORK

1041
00:41:34,463 --> 00:41:44,807
CLAUDIA, THANK YOU.

1042
00:41:45,941 --> 00:41:56,285
I THINK YOU'RE ON MUTE.

1043
00:42:11,333 --> 00:42:15,237
>> CAN YOU HEAR ME NOW IN
NYES

1044
00:42:15,237 --> 00:42:15,838
NGOOD AFTERNOON EVERYONE, THE

1045
00:42:15,838 --> 00:42:18,140
QUESTION IN OUR WILL BE IS TO

1046
00:42:18,140 --> 00:42:28,651
RECOGNIZE OW DNA IS DIAGRAMMED

1047
00:42:33,522 --> 00:42:35,758
AND PROCESSED.

1048
00:42:35,758 --> 00:42:38,460
--WHERE IT BINDS DNA POLYMERASES

1049
00:42:38,460 --> 00:42:40,863
AND IT PROCESSES DNA TO THE

1050
00:42:40,863 --> 00:42:41,730
REPLICATION REACTION EMPLOY

1051
00:42:41,730 --> 00:42:42,898
NONAPOPTOTIC YOU DNA LESIONS

1052
00:42:42,898 --> 00:42:44,867
SUCH AS THOSE INDUCED BIER UV

1053
00:42:44,867 --> 00:42:47,203
CAN BLOCK THE DNA POLYMERASE, SO

1054
00:42:47,203 --> 00:42:50,339
THIS CAN RESIDE IN DNA BREAKS

1055
00:42:50,339 --> 00:42:51,974
AND TRANSLOCATION, SO THE CELLS

1056
00:42:51,974 --> 00:42:53,475
ARE DEVELOPING FOR STAR

1057
00:42:53,475 --> 00:42:56,178
MECHANISM TO BYPASS THIS LESION.

1058
00:42:56,178 --> 00:43:00,149
IMPORTANT FOR OUR STUDY HAS BEEN

1059
00:43:00,149 --> 00:43:03,152
SHOWN THAT PC NA CAN BE POST

1060
00:43:03,152 --> 00:43:04,854
TRANSLATIONALLY MODIFIED FOR

1061
00:43:04,854 --> 00:43:07,923
WILL STALLS CAN GET

1062
00:43:07,923 --> 00:43:14,730
UBIQUITINNATEDDA THE LIGASE 164,

1063
00:43:14,730 --> 00:43:16,465
UBIQUITINNATED POLYMERASES ARE

1064
00:43:16,465 --> 00:43:19,702
ABLE TO BYPASS DNA LESIONS BUT

1065
00:43:19,702 --> 00:43:28,277
THEY CAN INTRODUCE MUTATIONS

1066
00:43:28,277 --> 00:43:28,978
ACROSS THE LESIONS.

1067
00:43:28,978 --> 00:43:31,747
SO WE WANTED TO FIND NEW FACTORS

1068
00:43:31,747 --> 00:43:33,349
THAT ARE INVOLVED IN THIS

1069
00:43:33,349 --> 00:43:35,017
PROCESS SO WE STARTED LOOKING

1070
00:43:35,017 --> 00:43:38,621
FOR PROTEINS THAT INTERACTED

1071
00:43:38,621 --> 00:43:39,688
WITH UBIQUITINNATED PUNISHING

1072
00:43:39,688 --> 00:43:39,855
CNA.

1073
00:43:39,855 --> 00:43:44,760
SO THIS IS HOW WE ENDED UP

1074
00:43:44,760 --> 00:43:45,895
STUDYING PAR10.

1075
00:43:45,895 --> 00:43:47,296
SO WHAT IS PAR10?

1076
00:43:47,296 --> 00:43:54,837
IT'S A MEMBER OF THE P A RP

1077
00:43:54,837 --> 00:43:56,605
FAMILY, AND AND THE SEQUENCE

1078
00:43:56,605 --> 00:44:00,042
STEPS, IN A MORE ADP ARRIVALS,

1079
00:44:00,042 --> 00:44:03,345
IN THE TRANSFER AND CHAINS THAT

1080
00:44:03,345 --> 00:44:07,683
CAN BE LINEAR OR BRANCHED.

1081
00:44:07,683 --> 00:44:11,086
THERE ARE 17--EVERYBODY TALKS

1082
00:44:11,086 --> 00:44:13,622
ABOUT PARP1 AND 2 BUT THERE ARE

1083
00:44:13,622 --> 00:44:15,758
17 MEMBERS IN THIS FAMILY, IT'S

1084
00:44:15,758 --> 00:44:17,226
TRUE THAT 1 AND 2 ARE THE MOST

1085
00:44:17,226 --> 00:44:19,795
STUDIED 1 ASK IT SHOWS THAT P A

1086
00:44:19,795 --> 00:44:22,665
RPs PLAY A ROLL IN SINGLE

1087
00:44:22,665 --> 00:44:30,739
STRADDED BREAKS AND FOR

1088
00:44:30,739 --> 00:44:31,807
OUR--IMPORTANT ROLE, THIS SUBSET

1089
00:44:31,807 --> 00:44:33,976
OF MEMBERS, THEY CAN ONLY

1090
00:44:33,976 --> 00:44:36,845
CATALYZE THE TRANSFER OF 1 ADP

1091
00:44:36,845 --> 00:44:38,547
RIEB OAZ, ON ACCEPTOR PROTEINS

1092
00:44:38,547 --> 00:44:40,616
THIS PROCESS IS CALLED

1093
00:44:40,616 --> 00:44:41,483
[INDISCERNIBLE] DPLI COSALATION

1094
00:44:41,483 --> 00:44:45,654
AND THIS SUBSET OF PROTEINS ARE

1095
00:44:45,654 --> 00:44:47,089
CALLED MONOATP RIBOSILL TRANSFER

1096
00:44:47,089 --> 00:44:50,526
ACES, SO P A RP10 IS A MEMBER OF

1097
00:44:50,526 --> 00:44:54,530
THIS SUBSET AND YOU CAN SEE IT

1098
00:44:54,530 --> 00:44:54,763
HERE.

1099
00:44:54,763 --> 00:44:57,132
NOW, WE DECIDED TO INVESTIGATE

1100
00:44:57,132 --> 00:45:02,404
ROLE OF P A RP10 IN DNA REPAIR

1101
00:45:02,404 --> 00:45:06,075
ABOUT YU P A R10, IT'S UNIQUE

1102
00:45:06,075 --> 00:45:09,378
WITHIN THE FAMILY BECAUSE IT HAS

1103
00:45:09,378 --> 00:45:11,280
A [INDISCERNIBLE] BOX

1104
00:45:11,280 --> 00:45:12,314
INTERACTING PEPTIDE AND WE START

1105
00:45:12,314 --> 00:45:14,984
AT FIRST WITH CONFIRMING THE

1106
00:45:14,984 --> 00:45:18,387
INTERACTION BETWEEN THE PC NA

1107
00:45:18,387 --> 00:45:20,589
AND PARP10 AND WE NEXT WANTED TO

1108
00:45:20,589 --> 00:45:22,691
KNOW WHAT'S THE ROLE OF THIS

1109
00:45:22,691 --> 00:45:25,494
INTERACTION IN AND SPECIFICALLY

1110
00:45:25,494 --> 00:45:27,930
IF IT'S ACTUALLY PARP10 REQUIRED

1111
00:45:27,930 --> 00:45:34,737
FOR REPLICATION RESTART THROUGH

1112
00:45:34,737 --> 00:45:35,004
TLS.

1113
00:45:35,004 --> 00:45:36,972
WE COULD SO THAT WHEN WE KNOCK

1114
00:45:36,972 --> 00:45:41,043
DOWN PARP10 WE SEE REDUCED PC NA

1115
00:45:41,043 --> 00:45:43,645
REDUCED UBIQUITINNATION, AND WE

1116
00:45:43,645 --> 00:45:46,815
USE SUBTLS TO SAY, AND WE SHOW

1117
00:45:46,815 --> 00:45:49,752
THAT YOU CAN USE THE MUTE

1118
00:45:49,752 --> 00:45:50,986
GENESIS IS USED AND KNOCKED

1119
00:45:50,986 --> 00:45:51,186
DOWN.

1120
00:45:51,186 --> 00:45:55,057
NOW THIS COULD BE RESTORE BIDE

1121
00:45:55,057 --> 00:45:57,659
REBRUCING WILD-TYPE PARP10 BUT

1122
00:45:57,659 --> 00:45:59,995
NOT MUTANT PARP10 THAT LACKS THE

1123
00:45:59,995 --> 00:46:05,434
INTERACTION WITH PC NA OR A

1124
00:46:05,434 --> 00:46:07,002
CATALYTIC ACTIVE MUTATION.

1125
00:46:07,002 --> 00:46:10,839
SO HOW DOES PARP10 PROMOTE TLS

1126
00:46:10,839 --> 00:46:14,443
IN WE THINK THAT BY REDUCING

1127
00:46:14,443 --> 00:46:15,644
UBIQUITINNATION, PARP10 IS

1128
00:46:15,644 --> 00:46:16,912
REDUCING THE POLYMERASES AND

1129
00:46:16,912 --> 00:46:23,786
WHAT WE SHOW, WE SHOW REDUCED

1130
00:46:23,786 --> 00:46:27,322
RAF 1 WHEN WE KNOCK DOWN PARP10

1131
00:46:27,322 --> 00:46:29,591
EMPLOY ACTUALLY CONFIRMING OUR

1132
00:46:29,591 --> 00:46:29,892
HYPOTHESIS.

1133
00:46:29,892 --> 00:46:32,428
SO WE HAVE THIS MODEL THAT

1134
00:46:32,428 --> 00:46:36,065
PARP10 IS RECRUITED TO

1135
00:46:36,065 --> 00:46:39,501
UBIQUITINNATED PC NA AND THROUGH

1136
00:46:39,501 --> 00:46:40,202
THIS CATALYTIC FUNCTION, FOR

1137
00:46:40,202 --> 00:46:42,738
MOST OF THE START OF THE

1138
00:46:42,738 --> 00:46:43,439
REPLICATION [INDISCERNIBLE]

1139
00:46:43,439 --> 00:46:49,478
THROUGH  TLS, THIS DATA HAS BEEN

1140
00:46:49,478 --> 00:46:51,480
PUBLISHED SOME TIME AGO AND I

1141
00:46:51,480 --> 00:46:54,783
WILL SHOW NEW DATA FROMMURE ON

1142
00:46:54,783 --> 00:46:55,017
LAB.

1143
00:46:55,017 --> 00:46:56,885
FOR SYNTHESIS, FORKS CAN BE

1144
00:46:56,885 --> 00:46:58,987
RESTARTED BOO I RIPRIMING AND

1145
00:46:58,987 --> 00:47:02,858
IT'S CATALYZED BY POL, BUT THIS

1146
00:47:02,858 --> 00:47:04,293
LEAVES BEHIND SINGLE STRAND DID,

1147
00:47:04,293 --> 00:47:04,660
NA GAPS.

1148
00:47:04,660 --> 00:47:07,296
THESE GAPS ARE TO BE FILLED AT

1149
00:47:07,296 --> 00:47:09,665
THE LATER STAGE.

1150
00:47:09,665 --> 00:47:12,101
NOW, IN THE LAST MAYBE 2 OR 3

1151
00:47:12,101 --> 00:47:15,170
YEARS, YEAH, THIS SINGLE STRAND

1152
00:47:15,170 --> 00:47:17,039
DNA GAPS ARE VERY, VERY

1153
00:47:17,039 --> 00:47:17,973
IMPORTANT BECAUSE IT HAS BEEN

1154
00:47:17,973 --> 00:47:20,142
SHOWN THAT THE ACCUMULATION OF

1155
00:47:20,142 --> 00:47:23,112
HE'S GAPS ACTUALLY CORRELATE

1156
00:47:23,112 --> 00:47:25,013
WITH SENSITIVITY OF CANCER CELLS

1157
00:47:25,013 --> 00:47:28,217
TO CHEMO THERAPY, PACIFICALLY IN

1158
00:47:28,217 --> 00:47:29,485
THE BRACKA DEFICIENT BACKGROUND.

1159
00:47:29,485 --> 00:47:33,188
SO HAS BEEN SHOWN THE LOSS OF

1160
00:47:33,188 --> 00:47:34,690
BRACCA PROTEINS CAUSES INCREASE

1161
00:47:34,690 --> 00:47:37,659
IN THE SINGLE STRAND FORMATION

1162
00:47:37,659 --> 00:47:40,162
THIS GAPS PERSIST AND THEY CAUSE

1163
00:47:40,162 --> 00:47:41,130
GENOMIC INABILITY AND CELL DEATH

1164
00:47:41,130 --> 00:47:51,373
AND WE CAN ACTUALLY EXPLOIT THAT

1165
00:47:51,373 --> 00:47:52,441
VULNERABILITY FOR THERAPY.

1166
00:47:52,441 --> 00:47:54,443
I SKIPPED 1 SLIDE.

1167
00:47:54,443 --> 00:47:57,312
YES, SO RECENT STUDY HAVE SHOWN

1168
00:47:57,312 --> 00:47:58,380
SEVERAL MECHANISMS FOR FILLING

1169
00:47:58,380 --> 00:48:01,350
UP THE SINGLE STRAND DNETWORKA

1170
00:48:01,350 --> 00:48:02,417
GAPS.

1171
00:48:02,417 --> 00:48:04,820
AND IN BRACCA DEFICIENT

1172
00:48:04,820 --> 00:48:05,921
BACKGROUND, THE SINGLE STRAND

1173
00:48:05,921 --> 00:48:08,323
DNA GAPS ARE FILLED OR REPAIRED

1174
00:48:08,323 --> 00:48:11,927
BY A TRANSLATION SYNTHESIS, AND

1175
00:48:11,927 --> 00:48:13,462
1 EXAMPLE IS [INDISCERNIBLE]

1176
00:48:13,462 --> 00:48:14,396
THAT HAS BEEN PUBLISH EXCLUDE

1177
00:48:14,396 --> 00:48:16,598
HAS BEEN SHOWN ALSO THAT

1178
00:48:16,598 --> 00:48:18,267
INHIBITION OF TLS PROTEINS IN

1179
00:48:18,267 --> 00:48:21,904
THE CELLS, CAUSED A ACCUMULATION

1180
00:48:21,904 --> 00:48:25,507
OF THE SINGLE STRAND GAPS.

1181
00:48:25,507 --> 00:48:28,977
NOW BECAUSE WE SHOWED PREVIOUSLY

1182
00:48:28,977 --> 00:48:30,379
THAT PARP10 PROMOTES

1183
00:48:30,379 --> 00:48:33,282
UBIQUITINNATION OF PC NA AND THE

1184
00:48:33,282 --> 00:48:34,783
PC NA UBIQUITINNATION HAS BEEN

1185
00:48:34,783 --> 00:48:36,318
SHOWN TO PLAY A ROLE IN GAP

1186
00:48:36,318 --> 00:48:38,053
FILLING WE WANT TO KNOW IF

1187
00:48:38,053 --> 00:48:40,255
PARP10 PLAYS A ROLE IN

1188
00:48:40,255 --> 00:48:41,390
GENERATING SINGLE STRAND DNA

1189
00:48:41,390 --> 00:48:47,596
GAPS SO FOR THIS WE USE BRD

1190
00:48:47,596 --> 00:48:47,829
COMMENT.

1191
00:48:47,829 --> 00:48:51,099
WE HAVE THE CELLS WITH THE BRDU,

1192
00:48:51,099 --> 00:48:52,968
WE COLLECT THEM, WE EMBED THEM,

1193
00:48:52,968 --> 00:48:57,873
WE PUT THEM ON SLIDES, WE RUN

1194
00:48:57,873 --> 00:49:00,175
WITH THE SINGLE CELL AND THEN WE

1195
00:49:00,175 --> 00:49:02,311
TAKE PICTURES OF THE BRDU

1196
00:49:02,311 --> 00:49:05,547
SIGNAL, SO WE CAN SHOW, AND WHEN

1197
00:49:05,547 --> 00:49:09,184
WE KNOCK DOWN PARP10, WE SEE

1198
00:49:09,184 --> 00:49:10,385
INCREASED SINGLE STRAND GAP

1199
00:49:10,385 --> 00:49:13,789
FORMATION, WHEN WE HAVE THE GAP

1200
00:49:13,789 --> 00:49:16,825
CONDITION, 0.4 MILLI MOLAR, THIS

1201
00:49:16,825 --> 00:49:20,028
HAS BEEN PREVIOUS LYE SHOWN IN

1202
00:49:20,028 --> 00:49:21,797
THE BRCCA BACKGROUND TO GENERATE

1203
00:49:21,797 --> 00:49:24,866
SINGLE STRAND DNA GAPS.

1204
00:49:24,866 --> 00:49:26,368
NOT EVERYBODY IN THE FIELD IS

1205
00:49:26,368 --> 00:49:29,938
USING THIS METHOD SO WE ACTUALLY

1206
00:49:29,938 --> 00:49:32,374
CONFIRMED THIS PHENOTYPE WITH S1

1207
00:49:32,374 --> 00:49:34,376
NUCLEACE DMA FIBER ASSAY, SO IN

1208
00:49:34,376 --> 00:49:37,746
THIS ASSAYS WE LABEL THE CELLS,

1209
00:49:37,746 --> 00:49:42,284
WE BIND THE ANALOGUES, WE

1210
00:49:42,284 --> 00:49:46,622
PROCESS THIS, TO OBTAIN LONG DNA

1211
00:49:46,622 --> 00:49:47,823
FIBERS THAT WE COME WITH A

1212
00:49:47,823 --> 00:49:49,524
SPECIAL MACHINE AND THEN WE

1213
00:49:49,524 --> 00:49:52,461
LABEL WITH SPECIFIC ANTIBODIES

1214
00:49:52,461 --> 00:49:55,464
FOR [INDISCERNIBLE], HALF THE

1215
00:49:55,464 --> 00:49:58,200
SAMPLES WE TREAT WITH THE S1

1216
00:49:58,200 --> 00:50:00,702
NUCLEACE, SO IF THE SINGLE

1217
00:50:00,702 --> 00:50:02,437
STRANDS ARE PRESCRIBINGENT AS 1

1218
00:50:02,437 --> 00:50:04,940
WOULD BELIEVE, THEN HAVE YOU A

1219
00:50:04,940 --> 00:50:06,742
SHORTER SECOND TRACK, A

1220
00:50:06,742 --> 00:50:07,476
[INDISCERNIBLE] TRACK EMPLOY SO

1221
00:50:07,476 --> 00:50:11,146
WHAT WE SEE IS WHEN WE KNOCK

1222
00:50:11,146 --> 00:50:13,348
DOWN PARP10, WE HAVE DECREASED

1223
00:50:13,348 --> 00:50:17,519
RATIO IN THE [INDISCERNIBLE]

1224
00:50:17,519 --> 00:50:19,821
DROP, WHEN WE TREAT WITH HU, SO

1225
00:50:19,821 --> 00:50:23,792
THIS IS A GAP INDUCED CONDITION.

1226
00:50:23,792 --> 00:50:25,460
BRACCA 2 KO WE USE THEM AS A

1227
00:50:25,460 --> 00:50:27,062
POSITIVE CONTROL.

1228
00:50:27,062 --> 00:50:29,197
WE ALSO GENERATED PARP10 AND WE

1229
00:50:29,197 --> 00:50:31,533
CAN CONFIRM THE SAME PHENOTYPE

1230
00:50:31,533 --> 00:50:34,036
IN THE PARP10 KNOCK OUT.

1231
00:50:34,036 --> 00:50:37,472
SO, WE CAN ACTUALLY SHOW THAT

1232
00:50:37,472 --> 00:50:39,107
PARP10 SUPPRESSES SINGLE STRAND

1233
00:50:39,107 --> 00:50:41,677
DNA GAP FORMATION, UPON

1234
00:50:41,677 --> 00:50:42,177
REPLICATION AND STRESS.

1235
00:50:42,177 --> 00:50:45,147
NEXT WE WANTED TO KNOW IF PARP10

1236
00:50:45,147 --> 00:50:51,820
IS ACTUALLY IN THE GAP FILLING

1237
00:50:51,820 --> 00:50:53,121
PROCESS, SO WE USE SERVE ASSAY,

1238
00:50:53,121 --> 00:50:55,657
THIS IS LIKE A [INDISCERNIBLE]

1239
00:50:55,657 --> 00:50:58,760
ASSAY, WE USE PARP10 ANTIBODY TO

1240
00:50:58,760 --> 00:51:02,030
SEE IF PARP10 LOCALIZES TO

1241
00:51:02,030 --> 00:51:04,032
NASCENT DNA UNDER THIS GAP

1242
00:51:04,032 --> 00:51:07,169
INDUCING CONDITIONS, SO WHAT WE

1243
00:51:07,169 --> 00:51:11,540
SEE IS WHEN WE TREAT CELLS, WE

1244
00:51:11,540 --> 00:51:13,308
ARE RECRUITMENT OF PARP10 TO THE

1245
00:51:13,308 --> 00:51:17,045
SINGLE STRAND DNA GAPS.

1246
00:51:17,045 --> 00:51:21,283
IT HAS BEEN SHOWN TO BE THE MAIN

1247
00:51:21,283 --> 00:51:22,150
UBIQUITIN LIGASE FOR PUNISHING

1248
00:51:22,150 --> 00:51:25,120
KRRK NA AND WE WANT TO KNOW FOR

1249
00:51:25,120 --> 00:51:26,388
18 AND PARP10 CO LOCALIZE IN

1250
00:51:26,388 --> 00:51:29,257
THESE CONDITION ANDS INDEED WE

1251
00:51:29,257 --> 00:51:32,394
SEE INCREASE FOCI 8 HOURS SIGNAL

1252
00:51:32,394 --> 00:51:35,764
WHEN WE CREATE EACH

1253
00:51:35,764 --> 00:51:36,131
[INDISCERNIBLE].

1254
00:51:36,131 --> 00:51:39,668
MORE IMPORTANT WHEN WE KNOCK

1255
00:51:39,668 --> 00:51:44,573
DOWN PARP10, WE HAVE A REDUCED

1256
00:51:44,573 --> 00:51:46,808
RECRUITMENT OF RAD 18 TO IN

1257
00:51:46,808 --> 00:51:50,712
NASCENT TREND GAPS SO THIS

1258
00:51:50,712 --> 00:51:52,481
ACTUALLY SUGGIESTS THAT PARP10

1259
00:51:52,481 --> 00:51:55,183
WAS RECRUITING RAD 18 TO THIS

1260
00:51:55,183 --> 00:51:55,384
SITE.

1261
00:51:55,384 --> 00:51:59,788
NEXT WE WANTED TO LOOK AT THE PC

1262
00:51:59,788 --> 00:52:03,191
NA UBIQUITINNATION LAB AND WE

1263
00:52:03,191 --> 00:52:04,259
USE THE UBIQUITINNATED PC NA

1264
00:52:04,259 --> 00:52:05,660
THAT IS NOW AVAILABLE AND WORKS

1265
00:52:05,660 --> 00:52:07,763
VERY GOOD SO WHEN WE KNOCK DOWN

1266
00:52:07,763 --> 00:52:15,170
PARP10, WE SEE REDUCED PC NA

1267
00:52:15,170 --> 00:52:16,905
UBIQUITINNATION, UNDER AND WE

1268
00:52:16,905 --> 00:52:20,876
ALSO RESERVE FOR 1 RECRUITMENT

1269
00:52:20,876 --> 00:52:22,844
WHEN WE KNOCK DOWN PARP10 IN

1270
00:52:22,844 --> 00:52:24,146
THIS 1 LOCATION, SO I SHOWED YOU

1271
00:52:24,146 --> 00:52:26,915
A LOT OF FIGURES BUT THE MESSAGE

1272
00:52:26,915 --> 00:52:31,286
IS, WE THINK PARP10 IS PROMOTING

1273
00:52:31,286 --> 00:52:32,087
MOTING

1274
00:52:32,087 --> 00:52:33,955
RECRUITMENT OF RAD 18 TO THESE

1275
00:52:33,955 --> 00:52:38,427
GAPS WHERE IT PROMOTES PC NA

1276
00:52:38,427 --> 00:52:40,662
UBIQUITINNATION AND RECRUITMENT

1277
00:52:40,662 --> 00:52:44,800
OF THE POLYMERASES.

1278
00:52:44,800 --> 00:52:45,534
WE GENERATEDINATENTS FOR PARP10

1279
00:52:45,534 --> 00:52:48,069
AND WE HAVE A CATALYTIC MUTANT

1280
00:52:48,069 --> 00:52:50,472
AND A BIG BOX MUTANT AND WE

1281
00:52:50,472 --> 00:52:52,340
ACTUALLY SHOWED THAT PARP10

1282
00:52:52,340 --> 00:52:53,642
REQUIRES THIS CATALYTIC ACTIVITY

1283
00:52:53,642 --> 00:52:57,279
BUT ALSO THE BCNA INTERACTION IN

1284
00:52:57,279 --> 00:53:03,018
ORDER TO DO GAP SUPPRESSION.

1285
00:53:03,018 --> 00:53:04,820
NOW AS HAS BEEN PREVIOUSLY SHOWN

1286
00:53:04,820 --> 00:53:07,989
THAT THE CELLS ACTUALLY RELY ON

1287
00:53:07,989 --> 00:53:10,258
TLS TO FIX THIS SINGLE STRAND

1288
00:53:10,258 --> 00:53:12,360
DNA GAPS SO WE WANTED TO KNOW IF

1289
00:53:12,360 --> 00:53:14,429
PARP10 PLAYS A ROLE IN THIS, SO

1290
00:53:14,429 --> 00:53:17,833
WHEN WE KNOCK DOWN PARP10 IN THE

1291
00:53:17,833 --> 00:53:20,836
BRARCA DEFESHT BACKGROUND, WE

1292
00:53:20,836 --> 00:53:22,404
SEE INCREED SINGLE STRAND GAP

1293
00:53:22,404 --> 00:53:24,272
FORMATION WHEN WE TREAT WITH HU

1294
00:53:24,272 --> 00:53:28,009
OR SIS PLATIN IS OUR GAP

1295
00:53:28,009 --> 00:53:30,045
INDUCING CONDITIONS.

1296
00:53:30,045 --> 00:53:33,815
WE ALSO ARE A PARP10 INHIBITOR

1297
00:53:33,815 --> 00:53:34,082
AVAILABLE.

1298
00:53:34,082 --> 00:53:35,283
PARP1 INHIBITORS DON'T WORK ON

1299
00:53:35,283 --> 00:53:38,386
PARP10 EMPLOY SO WELL IS PARP10

1300
00:53:38,386 --> 00:53:40,856
INHIBITOR AVAILABLE ASK WHEN WE

1301
00:53:40,856 --> 00:53:42,724
TREAT BRACCA DEFICIENT SIZE WITH

1302
00:53:42,724 --> 00:53:44,893
PARP10 INHIBITOR, WE ALSO SEE AN

1303
00:53:44,893 --> 00:53:51,600
INCREASE IN THE SINGLE STRAND

1304
00:53:51,600 --> 00:53:54,503
DNA GAPS, UNDER SOME INDUCING

1305
00:53:54,503 --> 00:53:54,936
CONDITIONS.

1306
00:53:54,936 --> 00:53:58,740
NOW, VERY IMPORTANT WHEN WE HAVE

1307
00:53:58,740 --> 00:54:00,442
BRACCA DEPLETED CELLS AND WE

1308
00:54:00,442 --> 00:54:02,811
TREAT THEM JUST ABOUT PARP10

1309
00:54:02,811 --> 00:54:04,880
INHIBITOR, WITHOUT ANY EXOGENOUS

1310
00:54:04,880 --> 00:54:10,318
DAMAGE, WE SEE REDUCED VIABILITY

1311
00:54:10,318 --> 00:54:16,558
IN BRACCA  CA AND THIS SUGGESTS

1312
00:54:16,558 --> 00:54:18,159
THAT THIS MAY ENHANCE THE

1313
00:54:18,159 --> 00:54:22,597
SENSITIVITY OF BECOME RAKRRK CA

1314
00:54:22,597 --> 00:54:23,999
DEFICIENT CELLS SOPHISTICATED

1315
00:54:23,999 --> 00:54:25,166
GENO TOXIC AGENTS SO THIS IS

1316
00:54:25,166 --> 00:54:26,268
SOMETHING WE WANT TO INVESTIGATE

1317
00:54:26,268 --> 00:54:26,935
IN THE FUTURE.

1318
00:54:26,935 --> 00:54:28,370
BECAUSE I ONLY HAD ABOUT 15

1319
00:54:28,370 --> 00:54:29,437
MINUTES TODAY AND I DIDN'T HAVE

1320
00:54:29,437 --> 00:54:30,972
TIME TO SHOW YOU EVERYTHING, I

1321
00:54:30,972 --> 00:54:32,274
JUST WANTED TO GIVE YOU AN

1322
00:54:32,274 --> 00:54:34,609
OVERVIEW OF WHAT WE ARE DOING

1323
00:54:34,609 --> 00:54:36,778
RIGHT NOW, I'M JUST GOING TO END

1324
00:54:36,778 --> 00:54:40,515
BY PROPOSING THIS MODEL, WE

1325
00:54:40,515 --> 00:54:44,386
THINK THAT PARP10, IT'S

1326
00:54:44,386 --> 00:54:45,186
PROMOTING GAP SUPPRESSION.

1327
00:54:45,186 --> 00:54:47,222
WE THINK THAT PARP10 DEPLETIONS

1328
00:54:47,222 --> 00:54:50,825
IMPAIRS THE ABILITY OF CELLS TO

1329
00:54:50,825 --> 00:54:52,427
REPAIR THIS GAPS USING

1330
00:54:52,427 --> 00:54:54,930
TRANSLATION SIPGHT SIS IN A

1331
00:54:54,930 --> 00:54:56,364
BRACCA PROFICIENT.

1332
00:54:56,364 --> 00:55:00,101
IF YOU HAVE BRACCA, PROFICIENT

1333
00:55:00,101 --> 00:55:01,937
CELLS, THESE CELLS ARE USING

1334
00:55:01,937 --> 00:55:03,338
HOMOLOGY BASE REPAIR TO REPAIR

1335
00:55:03,338 --> 00:55:09,511
THESE GAPS, BUT IN A BRACCA

1336
00:55:09,511 --> 00:55:12,480
DEFESHT BACKGROUND, THIS REPAIR

1337
00:55:12,480 --> 00:55:15,884
IS IMPAIRED, SO FURTHER LOSS OF

1338
00:55:15,884 --> 00:55:18,153
PARP10 REDUCES TRANSITION

1339
00:55:18,153 --> 00:55:18,954
SYNTHESIS GAP FILLING EMPLOY SO

1340
00:55:18,954 --> 00:55:20,922
THIS IS MY TAKE HOME MESSAGE FOR

1341
00:55:20,922 --> 00:55:21,189
YOU.

1342
00:55:21,189 --> 00:55:24,125
WE ARE STILL WORKING ON THIS.

1343
00:55:24,125 --> 00:55:25,660
I ONLY SHOW YOU PART OF THE

1344
00:55:25,660 --> 00:55:26,094
DATA.

1345
00:55:26,094 --> 00:55:29,164
WE HAVE THIS IN DIFFERENT CELL

1346
00:55:29,164 --> 00:55:31,166
LINES, WE CONFIRM IN DIFFERENT

1347
00:55:31,166 --> 00:55:36,404
SETTINGS, AND I'M GOING TO SAY

1348
00:55:36,404 --> 00:55:36,671
THANK YOU.

1349
00:55:36,671 --> 00:55:37,939
I WANT TO THANK A SPECIAL GROUP

1350
00:55:37,939 --> 00:55:42,177
OF PEOPLE THAT I WORK WITH,

1351
00:55:42,177 --> 00:55:43,912
YOU'D, ARBNAAND EMILY, AMAZING

1352
00:55:43,912 --> 00:55:45,180
PEOPLE AND [INDISCERNIBLE], TOO.

1353
00:55:45,180 --> 00:55:46,748
THIS IS OUR FUNDING FROM NETWORK

1354
00:55:46,748 --> 00:55:52,454
CI, AND I CAN TAKE QUESTIONS.

1355
00:55:52,454 --> 00:55:54,122
>> OKAY, CLAUDIA THAT WAS
REALLY

1356
00:55:54,122 --> 00:55:54,422
FANTASTIC.

1357
00:55:54,422 --> 00:55:56,191
WE ALREADY HAVE A BUNCH AS YOU

1358
00:55:56,191 --> 00:55:58,560
CAN IMAGINE OF QUESTIONS HERE IN

1359
00:55:58,560 --> 00:55:59,227
THE CHAT.

1360
00:55:59,227 --> 00:56:01,796
THE FIRST 2 FROM OUR PREVIOUS

1361
00:56:01,796 --> 00:56:03,131
SPEAKER ELISE, I WILL READ THEM

1362
00:56:03,131 --> 00:56:04,432
OFF 1 AT A TIME.

1363
00:56:04,432 --> 00:56:06,434
IN OUR COMMENT IN KOAMING

1364
00:56:06,434 --> 00:56:07,836
ASSAYS, HAVE YOU TESTED

1365
00:56:07,836 --> 00:56:12,073
REEXPRESSION OF A CATALYTIC

1366
00:56:12,073 --> 00:56:12,741
MUTANT OF PARP10?

1367
00:56:12,741 --> 00:56:19,714
>> YES, I THINK WE SHOWED
THAT.

1368
00:56:19,714 --> 00:56:23,852
YEAH, SO, WHEN WE REEXPRESS THE

1369
00:56:23,852 --> 00:56:26,354
CATALYTIC MUTANT, WE ACTUALLY

1370
00:56:26,354 --> 00:56:31,292
HAVE MORE GAPS, SO IF YOU PUT

1371
00:56:31,292 --> 00:56:32,627
BACK PARP10--

1372
00:56:32,627 --> 00:56:34,696
>> SORRY, I MISSED IT
THANK YOU.

1373
00:56:34,696 --> 00:56:35,764
>> AND THEN THE SECOND
QUESTION

1374
00:56:35,764 --> 00:56:38,933
IS ALSO FROM ELISE, HOW IT

1375
00:56:38,933 --> 00:56:40,602
PARP10 ACTIVATED, IS IT UPON

1376
00:56:40,602 --> 00:56:42,737
BINDING TO PC NA OR BIENING TO

1377
00:56:42,737 --> 00:56:44,939
DNA IN WHAT DO WE KNOW ABOUT IT?

1378
00:56:44,939 --> 00:56:46,474
NYES, WE ARE STILL LOOKING AT

1379
00:56:46,474 --> 00:56:46,675
THIS.

1380
00:56:46,675 --> 00:56:49,477
WE ARE STILL LOOKING AT THIS.

1381
00:56:49,477 --> 00:56:51,680
SO ACTUALLY PARP10 HAS AN

1382
00:56:51,680 --> 00:56:54,849
RRN DOMAIN, WE ARE LOOKING TO

1383
00:56:54,849 --> 00:56:57,385
SEE IF THIS RNN DOMAIN IS

1384
00:56:57,385 --> 00:56:58,920
IMPORTANT FOR BINDING OF PARP10

1385
00:56:58,920 --> 00:57:00,288
TO DNA, THIS IS OUR EXPERIMENTS

1386
00:57:00,288 --> 00:57:01,656
TO BE DONE IN THE FUTURE BUTOOSE

1387
00:57:01,656 --> 00:57:04,559
A VERY GOOD QUESTION.

1388
00:57:04,559 --> 00:57:07,162
NVERY INTERESTING, THANK YOU.

1389
00:57:07,162 --> 00:57:08,630
COMPREHEND NTHANK YOU SO MUCH.

1390
00:57:08,630 --> 00:57:10,799
>> ANOTHER QUESTION HERE
ALSO

1391
00:57:10,799 --> 00:57:16,738
FROM A FORMER PATTY CANNED 

1392
00:57:16,738 --> 00:57:17,539
CANNED--[INDISCERNIBLE] LAB

1393
00:57:17,539 --> 00:57:18,973
MEMBER SO IS RAD 18 CORRELATED

1394
00:57:18,973 --> 00:57:25,313
OR KNOWN TO BIND TO PAR OR

1395
00:57:25,313 --> 00:57:25,647
MARIN.

1396
00:57:25,647 --> 00:57:27,248
>> SO IT'S NOT PARALATION,
IT'S

1397
00:57:27,248 --> 00:57:31,886
MARALATION, SO WHEN YOU ADD IT'S

1398
00:57:31,886 --> 00:57:33,421
CALLED MONORIBOADP, SO WE HAVE

1399
00:57:33,421 --> 00:57:36,191
INVITRO ASSAYS WHERE WE DO

1400
00:57:36,191 --> 00:57:38,359
MODULATION, WE HAVE PRELIMINARY

1401
00:57:38,359 --> 00:57:39,994
DATA SHOWING THAT PARP10

1402
00:57:39,994 --> 00:57:41,796
MODULATES RAD 18, WE ARE TILL

1403
00:57:41,796 --> 00:57:44,332
CONFIRMING THAT ASSAYS, NOW THEY

1404
00:57:44,332 --> 00:57:48,670
DEVELOP NEW ANTIBODIES TO DETECT

1405
00:57:48,670 --> 00:57:49,537
MONOADPRICE O ISOLATION, WE ARE

1406
00:57:49,537 --> 00:57:53,141
TRYING THEM IN THE LAB, IT'S A

1407
00:57:53,141 --> 00:57:53,942
BIT OF TROUBLE TROUBLE SHOOTING

1408
00:57:53,942 --> 00:57:55,677
BUT THE DAILY BASIS THEA WAS NOT

1409
00:57:55,677 --> 00:57:56,745
READY FOR THIS PRESENTATION BUT

1410
00:57:56,745 --> 00:57:58,246
THANK YOU FOR THE QUESTION.

1411
00:57:58,246 --> 00:57:59,147
>> WE HAVE NO MORE
QUESTIONS

1412
00:57:59,147 --> 00:58:00,849
THERE IN THE CHAT BUT WE CAN

1413
00:58:00,849 --> 00:58:03,318
OPEN IT UP TO FURTHER QUESTIONS.

1414
00:58:03,318 --> 00:58:04,452
I HAVE A LOT OF QUESTIONS BUT

1415
00:58:04,452 --> 00:58:07,555
I'M GOING TO TALK TO YOU

1416
00:58:07,555 --> 00:58:09,591
AFTERWARDS.

1417
00:58:09,591 --> 00:58:19,834
>> PERFECT.

1418
00:58:21,102 --> 00:58:23,638
THANK YOU.

1419
00:58:23,638 --> 00:58:25,507
>> EVERYONE HAS TIGHT
SCHEDULES,

1420
00:58:25,507 --> 00:58:29,811
EVERYBODY WANTS TO BE DONE BY

1421
00:58:29,811 --> 00:58:30,044
1:30.

1422
00:58:30,044 --> 00:58:30,879
>> WE CAN KEEP GOING.

1423
00:58:30,879 --> 00:58:32,947
>> I HAVE MUCH MORE
DETAILED

1424
00:58:32,947 --> 00:58:34,382
DISCUSSIONS TO HAVE AND BOTHER

1425
00:58:34,382 --> 00:58:37,285
EVERYBODY ELSE BUT I THOUGHT IT

1426
00:58:37,285 --> 00:58:38,019
WAS REALLY FANTASTIC.

1427
00:58:38,019 --> 00:58:42,724
YOUR REVIEW THAT YOU WROTE FOR

1428
00:58:42,724 --> 00:58:44,392
CANCER WAS REALLY WONDERFUL AND

1429
00:58:44,392 --> 00:58:46,361
IT IS REALLY GREAT TO SEE THIS

1430
00:58:46,361 --> 00:58:46,728
MOVING FORWARD.

1431
00:58:46,728 --> 00:58:47,896
>> THANK YOU SO MUCH FOR

1432
00:58:47,896 --> 00:58:52,634
INVITING ME TO SPEAK TODAY.

1433
00:58:52,634 --> 00:58:52,901
N.

1434
00:58:52,901 --> 00:58:54,369
>> OKAY, IF THERE AREN'T
ANY

1435
00:58:54,369 --> 00:58:57,305
FURTHER QUESTIONS, I THINK IT

1436
00:58:57,305 --> 00:58:59,207
JUST LEAVES US TO THANK ALL THE

1437
00:58:59,207 --> 00:59:00,141
PRESENTERS TODAY FOR THE

1438
00:59:00,141 --> 00:59:01,676
EXCELLENT TALKS AND FOR KEEPING

1439
00:59:01,676 --> 00:59:02,477
ON TIME.

1440
00:59:02,477 --> 00:59:04,012
I'M PRETTY IMPRESSED AND ALSO TO

1441
00:59:04,012 --> 00:59:06,181
OUR ADVISORY BOARD MEMBERS FOR

1442
00:59:06,181 --> 00:59:07,048
THE COORDINATION OF THIS

1443
00:59:07,048 --> 00:59:07,482
WEBINAR.

1444
00:59:07,482 --> 00:59:10,018
WE WILL SEE YOU ALL NEXT MONTH

1445
00:59:10,018 --> 00:59:14,923
FOR A TALK BY ELEN LEEMAN,

1446
00:59:14,923 --> 00:59:16,024
ENTITLED TRANSLATION SYNTHESIS,

1447
00:59:16,024 --> 00:59:17,525
A DNA REVIEW AND

1448
00:59:17,525 --> 00:59:17,892
[INDISCERNIBLE].

1449
00:59:17,892 --> 00:59:19,494
WE HOPE TO SEE YOU ALL THEN AND

1450
00:59:19,494 --> 00:59:21,362
WITH THAT I WILL SAY GOODBYE TO

1451
00:59:21,362 --> 00:59:22,430
YOU ALL NTHANKS EVERYBODY.

1452
00:59:22,430 --> 00:59:24,165
GOOD TO SEE YOU.

1453
00:59:24,165 --> 00:59:24,632
THANK YOU.

1454
00:59:24,632 --> 00:59:25,567
 THANK YOU.

1455
00:59:25,567 --> 00:59:26,801
 THANKS EVERYONE.

1456
00:59:26,801 --> 00:59:37,078
THANK YOU.