1 00:00:05,280 --> 00:00:08,760 WELCOME TO THE JUNE EDITION OF 2 00:00:08,760 --> 00:00:10,560 THE NIH DNA REPAIR INTEREST 3 00:00:10,560 --> 00:00:14,200 GROUP VIDEO CONFERENCE SERIES. 4 00:00:14,200 --> 00:00:17,760 TODAY I WILL BE CO-HOSTING THE 5 00:00:17,760 --> 00:00:19,640 SEMINAR ALONG WITH KEN KRAMER 6 00:00:19,640 --> 00:00:23,440 AND OUR SPEAKER TODAY IS SHAN 7 00:00:23,440 --> 00:00:26,280 YAN FROM THE UNIVERSITY OF NORTH 8 00:00:26,280 --> 00:00:26,760 CAROLINA AT CHARLOTTE. 9 00:00:26,760 --> 00:00:27,720 BEFORE WE BEGENERATED I WANT TO 10 00:00:27,720 --> 00:00:30,080 ACKNOWLEDGE THE HELP WE GET WITH 11 00:00:30,080 --> 00:00:31,560 OUR SEMINAR SERIES FROM THE 12 00:00:31,560 --> 00:00:32,200 SCIENTIFIC ADVISORY BOARD WHO 13 00:00:32,200 --> 00:00:33,920 HELP US CHOOSE OUR SEMINAR 14 00:00:33,920 --> 00:00:37,040 SPEAKERS AND OUR IT PEOPLE 15 00:00:37,040 --> 00:00:38,320 INCLUDING FELICE HARPER WHO IS 16 00:00:38,320 --> 00:00:39,200 HELPING US WITH AV TODAY. 17 00:00:39,200 --> 00:00:41,160 AND I WANT TO REMIND YOU ALL 18 00:00:41,160 --> 00:00:43,560 THAT MANY PREVIOUS TALKS IN THIS 19 00:00:43,560 --> 00:00:46,760 SERIES CAN BE FOUND ON OUR VIDEO 20 00:00:46,760 --> 00:00:47,000 ARCHIVE. 21 00:00:47,000 --> 00:00:49,200 I ALSO WANT TO REMIND YOU THAT 22 00:00:49,200 --> 00:00:51,040 AFTER TODAY, THE WEBINARS WILL 23 00:00:51,040 --> 00:00:53,200 BE TAKING THE USUAL SUMMER BREAK 24 00:00:53,200 --> 00:00:55,920 NEAR JULY AND AUGUST AND WE WILL 25 00:00:55,920 --> 00:00:56,880 BE ROARING BACK IN SEPTEMBER 26 00:00:56,880 --> 00:00:59,840 WITH A TALK BY JOE MORRIS FROM 27 00:00:59,840 --> 00:01:01,080 THE UNIVERSITY OF BIRMINGHAM ON 28 00:01:01,080 --> 00:01:03,800 THE UK ON MECHANISMS OF 29 00:01:03,800 --> 00:01:04,360 SYNTHETIC LEATHALLITY. 30 00:01:04,360 --> 00:01:07,120 I ALSO WANTED TO TELL YOU THAT 31 00:01:07,120 --> 00:01:11,120 WE ARE SOLICITING NOMINATIONS 32 00:01:11,120 --> 00:01:14,160 FOR STELLAR POST DOCS AND JUNIOR 33 00:01:14,160 --> 00:01:15,720 FACULTY TO PRECEPT SHORT TALK 34 00:01:15,720 --> 00:01:17,600 NOTHING OUR ANNUAL SHOWCASE OF 35 00:01:17,600 --> 00:01:19,120 YOUNG INVESTIGATORS THAT WE WILL 36 00:01:19,120 --> 00:01:20,120 HEAD AS USUAL IN NOVEMBER OF 37 00:01:20,120 --> 00:01:21,080 THIS YEAR. 38 00:01:21,080 --> 00:01:22,160 SO NOMINATIONS SHOULD BE SENT TO 39 00:01:22,160 --> 00:01:23,400 MAY AS SOON AS POSSIBLE WITH THE 40 00:01:23,400 --> 00:01:25,600 NAME OF THE NOMINEE, THE CONTACT 41 00:01:25,600 --> 00:01:26,760 DETAILS AND A SHORT 42 00:01:26,760 --> 00:01:28,680 JUSTIFICATION FOR THE NOMINATION 43 00:01:28,680 --> 00:01:36,520 AND YOU CAN SEND THEM TO ME AT 44 00:01:36,520 --> 00:01:36,880 KARENU@NIH.GOV. 45 00:01:36,880 --> 00:01:40,520 NOW BACK TO TODAY'S SEMINAR, 46 00:01:40,520 --> 00:01:45,000 DR. SHAN YAN GOT HIS Ph.D. 47 00:01:45,000 --> 00:01:48,080 FROM THE UNIVERSITY, AND AFTER 48 00:01:48,080 --> 00:01:50,160 HARVARD JOINED THE FACULTY AT 49 00:01:50,160 --> 00:01:51,080 CHARLOTTE IN 2010. 50 00:01:51,080 --> 00:01:54,640 HE GOT TENURE IN 2016 AND BECAME 51 00:01:54,640 --> 00:01:59,320 FULL PROFESSOR IN 2019. 52 00:01:59,320 --> 00:02:01,120 SHAN DIRECTS THE BIOLOGY 53 00:02:01,120 --> 00:02:03,120 EXCHANGE GROUP THAT WAS ALSO AT 54 00:02:03,120 --> 00:02:05,080 THE BIOLOGY CENTER AND UNC CHAR 55 00:02:05,080 --> 00:02:06,120 LOT, ALSO ASSOCIATE CHAIR FOR 56 00:02:06,120 --> 00:02:08,880 RESEARCH IN THE DEPARTMENT OF 57 00:02:08,880 --> 00:02:09,680 BIOLOGICAL SCIENCES, AND PROGRAM 58 00:02:09,680 --> 00:02:12,520 LEAD IN THE GENOME INTEGRITY AND 59 00:02:12,520 --> 00:02:13,640 CANCER INITIATIVE AT UNC. 60 00:02:13,640 --> 00:02:17,560 HE IS GOING TO TELL US TODAY 61 00:02:17,560 --> 00:02:20,800 ABOUT THE ROLE OF ENDONUCLEACES 62 00:02:20,800 --> 00:02:21,920 IN PRESERVING GENOME INTEGRITY. 63 00:02:21,920 --> 00:02:25,560 BEFORE I HAND THINGS OVER TO 64 00:02:25,560 --> 00:02:27,040 SHAN, I WANT TO REMIND TO YOU 65 00:02:27,040 --> 00:02:29,000 PUT YOUR QUESTIONS TO HIM IN THE 66 00:02:29,000 --> 00:02:30,960 CHAT AND KEN AND I WILL READ 67 00:02:30,960 --> 00:02:32,960 THEM OUT AT THE END OF HIS TALK. 68 00:02:32,960 --> 00:02:33,440 SO I WILL STOPT SHARING MY 69 00:02:33,440 --> 00:02:35,520 SLIDES NOW AND HAND THE FLOOR TO 70 00:02:35,520 --> 00:02:36,600 SHAN. 71 00:02:36,600 --> 00:02:37,720 >> I WANT TO THANK 72 00:02:37,720 --> 00:02:41,040 THE HOSTS AND THE IT PERSON FOR 73 00:02:41,040 --> 00:02:43,240 HOSTING MY SEMINAR TODAY TO HAVE 74 00:02:43,240 --> 00:02:46,720 THIS OPPORTUNITY TO PRESENT WITH 75 00:02:46,720 --> 00:02:49,520 YOU OUR RECENT STUDY IN THE 76 00:02:49,520 --> 00:02:51,840 NUCLEUS 1 AND 2 IN GENOME 77 00:02:51,840 --> 00:02:54,720 INTEGRITY AND CANCER ETIOLOGY. 78 00:02:54,720 --> 00:02:56,680 THIS IS A SHORT OUTLINE, I WILL 79 00:02:56,680 --> 00:02:57,560 GIVE A SHORT INTRODUCTION FOR 80 00:02:57,560 --> 00:03:02,480 THOSE WHO ARE NOT FAMILIAR WITH 81 00:03:02,480 --> 00:03:06,680 APE2 AND 1 IN THE RESPONSE, THEN 82 00:03:06,680 --> 00:03:09,000 I WILL TALK ABOUT APE, AND THEN 83 00:03:09,000 --> 00:03:13,840 I WILL SWITCH TO APE1. 84 00:03:13,840 --> 00:03:17,480 SO [INDISCERNIBLE] CAN BE 85 00:03:17,480 --> 00:03:19,200 GENERATED BY SEVERAL EXOGENIUS 86 00:03:19,200 --> 00:03:22,400 SOURCES, ALL KINDS OF LEADERS 87 00:03:22,400 --> 00:03:26,280 AND IN PARTICULAR WE ARE 88 00:03:26,280 --> 00:03:32,560 INTERESTED IN APE-SIZE AND SSB 89 00:03:32,560 --> 00:03:35,360 TO CALCULATE HOW MANY PER DAY 90 00:03:35,360 --> 00:03:37,840 THERE ARE ROUGHLY 91 00:03:37,840 --> 00:03:40,800 [INDISCERNIBLE] APE SITE AND 92 00:03:40,800 --> 00:03:43,440 55,000 SSB SINGLE STRAND BREAKS 93 00:03:43,440 --> 00:03:48,080 EVEN IN NORMAL MAMMALIAN CELLS 94 00:03:48,080 --> 00:03:48,800 TODAY. 95 00:03:48,800 --> 00:03:50,840 TO REPAIR [INDISCERNIBLE] 96 00:03:50,840 --> 00:03:53,160 DAMAGE, DEPENDING ON THE EXACT 97 00:03:53,160 --> 00:03:55,960 NATURE OF DAMAGE, 98 00:03:55,960 --> 00:03:58,080 [INDISCERNIBLE] PATHWAYS CAN BE 99 00:03:58,080 --> 00:04:00,960 ACTIVELY REPAIR THEM INCLUDING 100 00:04:00,960 --> 00:04:03,000 BASIC EXCISION REPAIR, LET ME 101 00:04:03,000 --> 00:04:05,320 USE MY LASER POINTER HERE, 102 00:04:05,320 --> 00:04:11,600 SINGLE CELL REPAIR PATHWAY TO 103 00:04:11,600 --> 00:04:14,800 REPAIR SSB, AND OTHERS, THE 104 00:04:14,800 --> 00:04:19,600 HOMOLOGOUS COMBINATION AND 105 00:04:19,600 --> 00:04:22,040 COMPARE DSB, AND MMR, AND NER 106 00:04:22,040 --> 00:04:22,920 ARE ALSO COMPARE THEM. 107 00:04:22,920 --> 00:04:26,480 SO NEXT I WANT TO BRIEFLY 108 00:04:26,480 --> 00:04:27,480 INTRODUCE THIS BER PATHWAY TO 109 00:04:27,480 --> 00:04:29,360 YOU FOR THOSE THAT ARE NOT 110 00:04:29,360 --> 00:04:29,920 FAMILIAR WITH THIS. 111 00:04:29,920 --> 00:04:34,360 THE DAMAGE CAN BE RECOGNIZED AND 112 00:04:34,360 --> 00:04:36,560 PROCESSED BY [INDISCERNIBLE] TO 113 00:04:36,560 --> 00:04:38,560 GENERATE APE SIDE, BASICALLY NO 114 00:04:38,560 --> 00:04:43,240 BASICS, THEN THAT CAN BE FURTHER 115 00:04:43,240 --> 00:04:46,120 PROCESSED BY FUNCTIONAL GLUCOSE, 116 00:04:46,120 --> 00:04:49,480 OR APE1 [INDISCERNIBLE] AND THIS 117 00:04:49,480 --> 00:04:51,680 SINGLE STRAND BREAKS SSB CAN BE 118 00:04:51,680 --> 00:04:57,800 RECOGNIZED BY P A RP COMPLEXES 119 00:04:57,800 --> 00:04:59,040 AND XRCC1 AND FOLLOWING THAT 120 00:04:59,040 --> 00:05:01,480 THERE IS I BASIC EXCISION 121 00:05:01,480 --> 00:05:04,400 PATHWAY, THE DIFFERENCE IS 122 00:05:04,400 --> 00:05:08,000 [INDISCERNIBLE] IN WALL, 123 00:05:08,000 --> 00:05:11,760 [INDISCERNIBLE] MEDIATED 124 00:05:11,760 --> 00:05:12,040 PROCESSES. 125 00:05:12,040 --> 00:05:15,000 SINGLE STRAND BREAKS CAN BE 126 00:05:15,000 --> 00:05:17,200 DIFFERENT FROM DNA SUGAR DAMAGE, 127 00:05:17,200 --> 00:05:19,920 BASE DAMAGE AND A 128 00:05:19,920 --> 00:05:22,400 [INDISCERNIBLE] DAMAGE AND THAT 129 00:05:22,400 --> 00:05:23,960 CAN ARREST REPLICATION, 130 00:05:23,960 --> 00:05:25,160 TRANSCRIPTION PROGRAMS OR 131 00:05:25,160 --> 00:05:28,880 EXCESSIVE P A RP ACTIVATION 132 00:05:28,880 --> 00:05:30,880 LEADING TO GENOMICS ABILITY 133 00:05:30,880 --> 00:05:34,160 ESPECIALLY WITH CANCER, AND 134 00:05:34,160 --> 00:05:35,400 NEURODEGENERATIVE DISORDERS. 135 00:05:35,400 --> 00:05:37,920 THE SINGLE STRAND BREAKS CAN BE 136 00:05:37,920 --> 00:05:40,360 LOCALIZED IN NUCLEUS, ALSO 137 00:05:40,360 --> 00:05:43,080 RECENTLY FOUND IN MITOCHONDRIA 138 00:05:43,080 --> 00:05:44,560 AND EVEN IN CYTOPLASMS. 139 00:05:44,560 --> 00:05:47,440 NOW HOW TO REPAIR SINGLE STRAND 140 00:05:47,440 --> 00:05:52,080 BREAKS IS A CHALLENGE 141 00:05:52,080 --> 00:05:54,360 TECHNICALLY AND CONCEPTIONALLY 142 00:05:54,360 --> 00:05:57,040 BECAUSE THE BREAKING CAN CAUSE 143 00:05:57,040 --> 00:05:59,080 RAPID DISASSEMBLY O SO THEY CAN 144 00:05:59,080 --> 00:06:00,440 CAUSE PROBLEM REPLICATION EVEN 145 00:06:00,440 --> 00:06:03,480 WITH DEVELOPMENT OF TECHNICAL 146 00:06:03,480 --> 00:06:06,560 DEVELOPMENT FOR GENOME WIDE 147 00:06:06,560 --> 00:06:07,560 MAPPING OF DNA SINGLE STRAND 148 00:06:07,560 --> 00:06:11,000 BREAKS IN THE GENOME LIKE 149 00:06:11,000 --> 00:06:12,240 [INDISCERNIBLE] CAN MEASURE SOME 150 00:06:12,240 --> 00:06:15,000 TYPE OF SINGLE STRAND BREAKS BUT 151 00:06:15,000 --> 00:06:17,880 NOT ALL TYPES OF DNA SSB, SO 152 00:06:17,880 --> 00:06:21,560 IT'S WORTH IT TO STUDY THE 153 00:06:21,560 --> 00:06:25,800 SINGLE STRAND REPAIR AND 154 00:06:25,800 --> 00:06:26,400 SIGNALING PATHWAYS. 155 00:06:26,400 --> 00:06:29,720 FOR THOSE THAT ARE NOT FAMILIAR 156 00:06:29,720 --> 00:06:32,360 WITH THE DDR PATHWAYS THIS, IS A 157 00:06:32,360 --> 00:06:35,080 GOOD DIAGRAM TO SHOW THE DNA 158 00:06:35,080 --> 00:06:38,120 DAMAGE OR DNA STRESS CAN HANDLE 159 00:06:38,120 --> 00:06:40,680 SIGNALS AND THE SENSED LIKE 160 00:06:40,680 --> 00:06:42,160 MOLECULES LIKE OTHER SENSORS AND 161 00:06:42,160 --> 00:06:45,640 A SIGNAL NODE CAN BE TRANSDUCED 162 00:06:45,640 --> 00:06:48,840 TO DOWN STREAM TRANSDUCERS, 163 00:06:48,840 --> 00:06:51,000 EFFECTORS, EVENTUALLY TO ARREST 164 00:06:51,000 --> 00:06:54,880 CANNED WHAT CAUSE APOPTOSIS 165 00:06:54,880 --> 00:06:57,000 SENESCENSEL, AFTER TRANSCRIPTION 166 00:06:57,000 --> 00:06:58,120 AND REPAIR DNA DAMAGE. 167 00:06:58,120 --> 00:07:00,560 IN RESPONSE TO THOSE,S ARE 168 00:07:00,560 --> 00:07:03,040 ESPECIALLY DOUBLE STRAND BREAKS, 169 00:07:03,040 --> 00:07:07,760 2 KEY PI3 KINASES, ATM AND 170 00:07:07,760 --> 00:07:11,920 [INDISCERNIBLE] TO PREPARE FOR 171 00:07:11,920 --> 00:07:13,560 REPAIR SIGNALING FOR 172 00:07:13,560 --> 00:07:14,920 [INDISCERNIBLE], THE SINGLE 173 00:07:14,920 --> 00:07:18,400 STRAND DNA CAN CODED WITH RPA, 174 00:07:18,400 --> 00:07:21,880 THAT CAN TRIGGER ATR KINASE 175 00:07:21,880 --> 00:07:25,760 ACTIVATION TOGETHER WITH ATR 176 00:07:25,760 --> 00:07:28,000 INTACT PROTEIN COMPLEXES. 177 00:07:28,000 --> 00:07:30,840 THE DDR PATHWAY IS VERY COMPLEX, 178 00:07:30,840 --> 00:07:36,000 BACK IN 1997, WE ONLY KNOW THE 179 00:07:36,000 --> 00:07:39,080 GENE DAMAGE BETWEEN ATRIP, THAT 180 00:07:39,080 --> 00:07:40,840 IS P3 ACTIVATED AND THE ACTIVITY 181 00:07:40,840 --> 00:07:43,640 CAUSE THE CYCLE ARREST, LATER IN 182 00:07:43,640 --> 00:07:47,480 ON IN 2007 IT'S ALREADY SO 183 00:07:47,480 --> 00:07:49,440 COMPLEX, DEPENDING ON THE TYPE 184 00:07:49,440 --> 00:07:52,160 OF [INDISCERNIBLE], THE POSSIBLE 185 00:07:52,160 --> 00:07:54,600 CHOICE TO TRIGGER DDR PATHWAYS. 186 00:07:54,600 --> 00:07:59,040 IN 2017 THIS NICE REVIEW FROM 187 00:07:59,040 --> 00:08:00,960 [INDISCERNIBLE] SUMMARIZED HOW 188 00:08:00,960 --> 00:08:04,240 THIS ATR DDR PATHWAY IS 189 00:08:04,240 --> 00:08:05,400 ACTIVATED. 190 00:08:05,400 --> 00:08:08,200 BRIEFLY IN RESPONSE TO 191 00:08:08,200 --> 00:08:09,600 [INDISCERNIBLE] SINGLE STRAND 192 00:08:09,600 --> 00:08:12,040 BREAK, SINGLE STRAND DNA IS 193 00:08:12,040 --> 00:08:12,840 GENERATED, THE [INDISCERNIBLE] 194 00:08:12,840 --> 00:08:16,680 WILL BE CODED AND THE ATR IS 195 00:08:16,680 --> 00:08:18,320 INTACT [INDISCERNIBLE] COMPLEX 196 00:08:18,320 --> 00:08:20,880 CAN BE RECORDED TO RPA CODED 197 00:08:20,880 --> 00:08:23,600 SINGLE STRAND DNA, THROUGH THE 198 00:08:23,600 --> 00:08:26,120 TECHNICALLY TO AFIB AND RPA. 199 00:08:26,120 --> 00:08:28,720 THEY'RE 2 DIRECT ACTIVATOR 200 00:08:28,720 --> 00:08:33,440 PROTEIN OF ATR CALLED TOP AP1 201 00:08:33,440 --> 00:08:36,320 AND ETWAY 1, BOTH APPROACH WILL 202 00:08:36,320 --> 00:08:39,000 BE RECRUITED TO THAT RP-CODED 203 00:08:39,000 --> 00:08:41,400 SINGLE STRAND DNA SO THAT ATR 204 00:08:41,400 --> 00:08:46,160 KINASE WILL BE ACTIVATED, SO, SO 205 00:08:46,160 --> 00:08:51,120 FAR, TOP 1 AND EETAC1 ARE THE 2 206 00:08:51,120 --> 00:08:53,880 MOST WIDELY RECOGNIZED DIRECT 207 00:08:53,880 --> 00:08:55,880 ACTIVATOR MOLECULES OF ATR 208 00:08:55,880 --> 00:08:57,320 KINASES, WE WILL COME BACK TO 209 00:08:57,320 --> 00:08:58,000 THAT LATER. 210 00:08:58,000 --> 00:09:02,360 SO WHY DO I STUDY ATR/DDR 211 00:09:02,360 --> 00:09:02,640 PATHWAY. 212 00:09:02,640 --> 00:09:04,360 NOW 10 YEARS AGO, MORE THAN 10 213 00:09:04,360 --> 00:09:07,720 YEARS AGO, THERE'S A PAPER SHOWS 214 00:09:07,720 --> 00:09:15,280 THE FIRST ATR INHIBITOR, 215 00:09:15,280 --> 00:09:16,640 [INDISCERNIBLE] CAN SELECTIVELY 216 00:09:16,640 --> 00:09:19,840 KILL CANCER CELLS AND IN 217 00:09:19,840 --> 00:09:20,920 CLINICAL TRIALS THE FIRST 218 00:09:20,920 --> 00:09:24,640 REPORTER CAME IN 3 YEARS AGO, 219 00:09:24,640 --> 00:09:27,400 THE PHASE 1 CLINICAL TRIALS KIND 220 00:09:27,400 --> 00:09:33,520 OF A MORE DEVELOPED BURDEN, 221 00:09:33,520 --> 00:09:35,200 TARGETING KINASES, WITH VERY 222 00:09:35,200 --> 00:09:41,400 ENCOURAGING NEWS ON TARGETING 223 00:09:41,400 --> 00:09:42,960 ATR DDR FOR CANCER THERAPEUTICS. 224 00:09:42,960 --> 00:09:45,120 THERE ARE ALSO OTHERS AT 225 00:09:45,120 --> 00:09:47,080 DIFFERENT STATUS OF CLINICAL 226 00:09:47,080 --> 00:09:47,440 TRIALS. 227 00:09:47,440 --> 00:09:50,920 NOW I WANT TO INTRODUCE SOME 228 00:09:50,920 --> 00:09:53,040 BACKGROUND OF MY 2 FAVORITE 229 00:09:53,040 --> 00:09:58,920 PROTEIN, APE 1 AND APE 2. 230 00:09:58,920 --> 00:10:00,560 SO DR. [INDISCERNIBLE] AND 231 00:10:00,560 --> 00:10:02,840 DR. WILSON POSTED THIS REVIEW ON 232 00:10:02,840 --> 00:10:05,520 APE1, SO THIS IS PRESCRIBING 233 00:10:05,520 --> 00:10:08,320 NEBT APENUCLEUS 1, IT HAS END 234 00:10:08,320 --> 00:10:08,680 TERMINAL DOMAIN, 235 00:10:08,680 --> 00:10:11,360 [INDISCERNIBLE], IT AWLINGS ARE 236 00:10:11,360 --> 00:10:16,800 HAS ASSISTANCE UP TO 5 REMEDIATE 237 00:10:16,800 --> 00:10:22,120 REDOX ACTIVITY AND AS THE 238 00:10:22,120 --> 00:10:22,920 ENDONUCLEUS, PHOSPHOR 239 00:10:22,920 --> 00:10:24,840 DISCIPLINARY FORUS MEDIATED 240 00:10:24,840 --> 00:10:27,920 ACTIVITY, SO IT'S MAINLY 241 00:10:27,920 --> 00:10:29,560 FUNCTION IN THE PAIR, IN 242 00:10:29,560 --> 00:10:31,600 PROCESSING AND TRANSCRIPTION OF 243 00:10:31,600 --> 00:10:34,080 APE1 FROM LITERATURE. 244 00:10:34,080 --> 00:10:37,120 HOWEVER, IF YOU LOOK AT 245 00:10:37,120 --> 00:10:39,560 LITERATURE, TARGETING APE 1 IN 246 00:10:39,560 --> 00:10:41,960 THE TREATMENT OF HUMAN DISEASES, 247 00:10:41,960 --> 00:10:45,400 THERE'S ONLY 1 IN CLINICAL 248 00:10:45,400 --> 00:10:47,160 TRIALS. 249 00:10:47,160 --> 00:10:50,760 APX 3330 TARGETING IS REDOX 250 00:10:50,760 --> 00:10:51,800 FUNCTION, EVEN THOUGH--IT'S WELL 251 00:10:51,800 --> 00:10:54,080 KNOWN FOR FUNCTION IN DEAN 252 00:10:54,080 --> 00:10:58,040 REPAIR, BUT IT'S MECHANISM FOR 253 00:10:58,040 --> 00:10:59,920 OUR METABOLISM SINCE THE PLOT 254 00:10:59,920 --> 00:11:01,720 INDUCED THE [INDISCERNIBLE] AND 255 00:11:01,720 --> 00:11:04,400 GENE DAMAGE RESPONSE IS NOT VERY 256 00:11:04,400 --> 00:11:04,600 CLEAR. 257 00:11:04,600 --> 00:11:07,680 EVEN THOUGH WE HAVE PUBLISHED 258 00:11:07,680 --> 00:11:11,360 LIKE 1500 PAPERS, ON APE1 IN PUB 259 00:11:11,360 --> 00:11:15,360 MED SO THERE'S A LOT OF 260 00:11:15,360 --> 00:11:16,160 [INDISCERNIBLE] MECHANISM FOR 261 00:11:16,160 --> 00:11:16,360 APE1. 262 00:11:16,360 --> 00:11:21,960 NOW I WILL MOVE ON TO APE2. 263 00:11:21,960 --> 00:11:22,840 APEINTERNUCLEUS 2, IN MANY 264 00:11:22,840 --> 00:11:25,400 DIFFERENT SPECS MAY HAVE 265 00:11:25,400 --> 00:11:29,040 DIFFERENT NAME, CALLED MPN2, OR 266 00:11:29,040 --> 00:11:31,440 ONLY LIKE 70-80 PAPERS IN PUB 267 00:11:31,440 --> 00:11:32,800 MED SO FAR. 268 00:11:32,800 --> 00:11:34,800 IT ALSO HAS EEP AT THE END 269 00:11:34,800 --> 00:11:35,840 TERMINUS, AND AT THE SAME 270 00:11:35,840 --> 00:11:37,520 TERMINUS IT HAS OTHER 2 FACTOR 271 00:11:37,520 --> 00:11:41,520 DOMAIN, 1 IS CALLED PAPER 272 00:11:41,520 --> 00:11:42,960 [INDISCERNIBLE] MEDIATED PC 273 00:11:42,960 --> 00:11:45,080 INTEX, FOR THE MOTIF. 274 00:11:45,080 --> 00:11:49,360 ALL THIS 3 FUNCTION MOTIFS ARE 275 00:11:49,360 --> 00:11:50,800 CONSERVED DURING EVOLUTION, FROM 276 00:11:50,800 --> 00:11:53,000 DIFFERENT SPECIES OF APE TO 277 00:11:53,000 --> 00:11:54,360 [INDISCERNIBLE], AND WE 278 00:11:54,360 --> 00:11:56,880 SUMMARIZE THIS APE TO 279 00:11:56,880 --> 00:11:59,520 FUNCTIONALITIES IN GENOME AND 280 00:11:59,520 --> 00:12:02,200 EPIGENOME INTEGRITY MAINLY IN 281 00:12:02,200 --> 00:12:05,440 DNA REPAIR, IMMUNE RESPONSE 282 00:12:05,440 --> 00:12:11,400 EPIGENETIC REGULATION. 283 00:12:11,400 --> 00:12:21,880 OR RECENT STUDY FUNCTION--I 284 00:12:24,880 --> 00:12:27,080 THINK 4 OR 5 STUDIES HILIGHTS 285 00:12:27,080 --> 00:12:28,120 THE APE 1 OR 2. 286 00:12:28,120 --> 00:12:30,360 THIS IS THE FIRST WOIN CAME TBR 287 00:12:30,360 --> 00:12:33,920 STEVE'S LAB AT HARVARD MEDICAL 288 00:12:33,920 --> 00:12:41,120 SCHOOL USING CRSPR SCREENING OF 289 00:12:41,120 --> 00:12:42,680 GENOME WIDE BRCA 2 DEFICIENT 290 00:12:42,680 --> 00:12:44,120 CANCER CELLS. 291 00:12:44,120 --> 00:12:49,360 IT CAME WITH APE 2 AS THE MOST 292 00:12:49,360 --> 00:12:52,120 PROFOUND SYNTHETIC LETHAL FOR 293 00:12:52,120 --> 00:12:56,880 BRCA2 DEFICIENCY AND PROPOSED 294 00:12:56,880 --> 00:12:57,560 APEFUNCTION IN EXCISION REPAIR 295 00:12:57,560 --> 00:13:08,000 MAY BE THE REASON FOR THE 296 00:13:23,560 --> 00:13:24,000 [INDISCERNIBLE]. 297 00:13:24,000 --> 00:13:25,720 THEN WILLIAMS PUT TOGETHER A 298 00:13:25,720 --> 00:13:29,320 PAPER NOT ONLY FOR BRCA 299 00:13:29,320 --> 00:13:32,000 DEFICIENT BUT BRCAC1 CELLS, APE2 300 00:13:32,000 --> 00:13:34,920 IS THE EFFECTIVE LETHAL TARGET 301 00:13:34,920 --> 00:13:41,200 AND ALSO PROPOSE APE DYSFUNCTION 302 00:13:41,200 --> 00:13:44,240 IN THE RESOLUTION OF 303 00:13:44,240 --> 00:13:46,120 [INDISCERNIBLE]. 304 00:13:46,120 --> 00:13:48,840 THIS YEAR, FROM 2003, 305 00:13:48,840 --> 00:13:49,760 [INDISCERNIBLE]'S LAB PUBLISHED 306 00:13:49,760 --> 00:13:51,840 THIS CELL PAPER REPORTING THAT 307 00:13:51,840 --> 00:13:57,720 THE FUNCTION OF APE2 IN THE MMEG 308 00:13:57,720 --> 00:13:59,320 R J ACTIVITY ESPECIALLY IN THE 309 00:13:59,320 --> 00:14:02,080 RESOLUTION OF THE 3 PRIME OF 3 310 00:14:02,080 --> 00:14:06,160 FLAT STRUCTURES THROUGH IT'S EEP 311 00:14:06,160 --> 00:14:06,880 DOMAIN. 312 00:14:06,880 --> 00:14:09,840 SO ALL THOSE GENOME WIDE STRING 313 00:14:09,840 --> 00:14:11,440 POINTING OUT THAT THE 314 00:14:11,440 --> 00:14:16,280 SIGNIFICANCE OR APE1 OR 2, AS A 315 00:14:16,280 --> 00:14:22,040 SYNTHETIC TARGET BRCA 1 AND 2 316 00:14:22,040 --> 00:14:22,600 DEFICIENT CELLS. 317 00:14:22,600 --> 00:14:25,280 SO IF YOU LOOK AT APE1 AND 2 318 00:14:25,280 --> 00:14:30,320 STRUCTURE, THEY ALL SHARED THE 319 00:14:30,320 --> 00:14:34,360 EEP DOMINANT, THE END TERMINAL 320 00:14:34,360 --> 00:14:37,000 HAS KRAMER FRAGMENT AS 321 00:14:37,000 --> 00:14:39,760 MISINTERPRETED DOMAIN, REDOX 322 00:14:39,760 --> 00:14:41,240 [INDISCERNIBLE] AND EXTRA DOMAIN 323 00:14:41,240 --> 00:14:44,280 AS C-TERMINUS, IF YOU LOOK AT 324 00:14:44,280 --> 00:14:46,360 THE ACTIVITY, APE1 HAS VERY 325 00:14:46,360 --> 00:14:50,320 STRONG OR FAST APEIN THE NUCLEUS 326 00:14:50,320 --> 00:14:53,480 BUT WERE WE GO SLOW 327 00:14:53,480 --> 00:14:54,160 [INDISCERNIBLE] ACTIVITY, APE 328 00:14:54,160 --> 00:14:56,880 WILL KNOCK OUT THE BRCA LETHAL 329 00:14:56,880 --> 00:14:59,200 AND APE2 AND OPPOSITE, IT HAS 330 00:14:59,200 --> 00:15:01,960 WEAK OR SLOW APE IN THE NUCLEIC 331 00:15:01,960 --> 00:15:06,520 ACTIVITY BUT A STRONG OR FIRST 332 00:15:06,520 --> 00:15:07,920 NUCLEIC ACTIVITY. 333 00:15:07,920 --> 00:15:11,760 THE APE2 KNOCK OUT IS NOT LETHAL 334 00:15:11,760 --> 00:15:13,960 BUT HAS SOME DEFICIENCY IN THE 335 00:15:13,960 --> 00:15:14,240 CELL CYCLE. 336 00:15:14,240 --> 00:15:18,600 SO WHEN I SEARCH A WEBSITE, I 337 00:15:18,600 --> 00:15:21,600 SAW THIS APE1, AND OLDER BROTHER 338 00:15:21,600 --> 00:15:24,040 AND YOUNGER BROTHER AND APE1 MAY 339 00:15:24,040 --> 00:15:25,880 SAY OH BROTHER, I WAS DISCOVERED 340 00:15:25,880 --> 00:15:27,200 FIRST SO I CAN DO EVERYTHING 341 00:15:27,200 --> 00:15:30,840 BECAUSE IF YOU CAN SEE, YOU CAN 342 00:15:30,840 --> 00:15:33,400 FIND 1500 PAPERS IN PUB MED, BUT 343 00:15:33,400 --> 00:15:35,120 APE2 MAY SAY OH REALLY, MAYBE I 344 00:15:35,120 --> 00:15:36,760 CAN SEE THAT YOU DON'T EVEN 345 00:15:36,760 --> 00:15:38,520 THOUGH THERE ARE ONLY 70 PAPERS 346 00:15:38,520 --> 00:15:39,040 IN PUB MED. 347 00:15:39,040 --> 00:15:46,600 SO TO FIGURE OUT WHAT APE1 TO DO 348 00:15:46,600 --> 00:15:48,440 TO [INDISCERNIBLE] STABILITY IN 349 00:15:48,440 --> 00:15:49,960 CANCER, WE PRIMARILY USE THIS AS 350 00:15:49,960 --> 00:15:51,480 A MODEL SYSTEM FOR THOSE WHO ARE 351 00:15:51,480 --> 00:15:52,880 NOT FAMILIAR WITH THE SYSTEM, 352 00:15:52,880 --> 00:16:03,440 THIS IS A BRIEF INTRODUCTION SO 353 00:16:04,760 --> 00:16:07,400 XENOPUS, IT'S LOW SPEED 354 00:16:07,400 --> 00:16:08,440 SUPERINATE BUT WITHOUT 355 00:16:08,440 --> 00:16:08,880 RESOURCES. 356 00:16:08,880 --> 00:16:12,720 IF YOU USE HIGHER SPEED, YOU CAN 357 00:16:12,720 --> 00:16:15,600 GET A LAYER OF THE MEMBRANE AND 358 00:16:15,600 --> 00:16:19,840 WE WILL CARVE THIS HSS. 359 00:16:19,840 --> 00:16:22,160 AND YOU ADD A [INDISCERNIBLE] 360 00:16:22,160 --> 00:16:26,440 DNA [INDISCERNIBLE] THAT CAN 361 00:16:26,440 --> 00:16:29,200 FORM A NUCLEAR MEMBRANE, IT CAN 362 00:16:29,200 --> 00:16:31,920 BE REPLICATED, YOU CAN ALSO 363 00:16:31,920 --> 00:16:34,040 ICEOSEALATE THOSE NCLUE I AND 364 00:16:34,040 --> 00:16:35,840 THEN GET NUCLEAR LAYER, CAN YOU 365 00:16:35,840 --> 00:16:43,200 FURTHER SPIN DOWN, YOU CAN GET 366 00:16:43,200 --> 00:16:44,520 MIMIC MP EXTRACTS. 367 00:16:44,520 --> 00:16:48,400 SO LSS, AND HSS, CAN BE USED FOR 368 00:16:48,400 --> 00:16:52,840 STUDY GENE PAIR AND DDR PATH 369 00:16:52,840 --> 00:16:55,480 BASE BECAUSE LSSS, YOU ADD 370 00:16:55,480 --> 00:16:58,880 NUCLEI, AND YOU CAN DAMAGE THE 371 00:16:58,880 --> 00:17:01,680 [INDISCERNIBLE] WITH OTHER GENE 372 00:17:01,680 --> 00:17:04,240 DAMAGING AGENTS CAN ALSO DEPLETE 373 00:17:04,240 --> 00:17:06,120 A TARGETED PROTEIN, OR THE 374 00:17:06,120 --> 00:17:10,400 WILD-TYPE OR MUTANT PROTEIN, CAN 375 00:17:10,400 --> 00:17:11,680 YOU REPAIR OR OTHER THING. 376 00:17:11,680 --> 00:17:15,400 THE GOOD THING FOR HSS, IS YOU 377 00:17:15,400 --> 00:17:17,760 CAN ADD DEFINED STRUCTURES LIKE 378 00:17:17,760 --> 00:17:20,240 PLAZ MIDS MIDDNA WITH THE 379 00:17:20,240 --> 00:17:21,080 DEFINED GENE [INDISCERNIBLE] 380 00:17:21,080 --> 00:17:24,200 THAT YOU CAN LOCATE THE GENE 381 00:17:24,200 --> 00:17:26,200 REPAIR DYNAMICS, DDR PATHWAYS 382 00:17:26,200 --> 00:17:26,520 ACTIVATIONS. 383 00:17:26,520 --> 00:17:28,960 SO LET'S TALK ABOUT THE FUNCTION 384 00:17:28,960 --> 00:17:35,320 OF APE 2 IN ATR [INDISCERNIBLE] 385 00:17:35,320 --> 00:17:38,640 IN CANCER BIOLOGY .D BACK IN 386 00:17:38,640 --> 00:17:41,040 2013 MY GRADUATE STUDENT JEREMY 387 00:17:41,040 --> 00:17:44,560 TO HONORS GRADUATE STUDENTS 388 00:17:44,560 --> 00:17:46,480 [INDISCERNIBLE] TO STUDY WHAT'S 389 00:17:46,480 --> 00:17:48,120 THE FUNCTIONAL APCEREBELLUMS 2 390 00:17:48,120 --> 00:17:50,920 IN HYDROGEN PEROXIDE INDUCE SAYS 391 00:17:50,920 --> 00:17:53,480 ATR PATHWAY USING XENOPUSAS A 392 00:17:53,480 --> 00:17:54,720 SYSTEM. 393 00:17:54,720 --> 00:17:58,600 SO PRIMARILY WE USE H22 TO 394 00:17:58,600 --> 00:18:00,880 DAMAGE CHROMATIN DNA TO CHECK 395 00:18:00,880 --> 00:18:07,400 FOR PHOSPHORYLATION WHICH IS 396 00:18:07,400 --> 00:18:08,880 INDICATOR ISOLATION, WE FOUND 397 00:18:08,880 --> 00:18:12,360 IT'S LOADED TO THE KROEMA TIN 398 00:18:12,360 --> 00:18:16,160 DNA OR TO THE COMPLEXES AND THE 399 00:18:16,160 --> 00:18:21,200 RETINA AS NORMAL COMPLEXES, ALSO 400 00:18:21,200 --> 00:18:23,920 HIGHLY ENRICHED CHROMATIN DNA. 401 00:18:23,920 --> 00:18:26,520 WHEN WE USE DEPLETION, TARGETING 402 00:18:26,520 --> 00:18:31,720 APE 2, IN THE NAP2'S GONE, THE 403 00:18:31,720 --> 00:18:32,200 PHOSPHORYLATION BY IT'S 404 00:18:32,200 --> 00:18:35,840 COMPROMISED, ALL THE ATR, 405 00:18:35,840 --> 00:18:39,680 COMPLEXES, RECRUITMENT IS 406 00:18:39,680 --> 00:18:41,520 COMPROMISED, WHERE WE MAKE AIT 407 00:18:41,520 --> 00:18:44,240 PE RECOMBIN ANT PROTEINS AND WE 408 00:18:44,240 --> 00:18:46,120 FIND THE PHOSPHORYLATION BY H2 409 00:18:46,120 --> 00:18:49,400 IS RESCUED, THE RECRUITMENT OF 410 00:18:49,400 --> 00:18:51,440 APE KROM COMPLEX TO DAMAGED 411 00:18:51,440 --> 00:18:52,880 PROTEINS IS ALSO SKEWED. 412 00:18:52,880 --> 00:18:57,000 SO THAT MEANS APE2 IS CRITICAL 413 00:18:57,000 --> 00:19:00,760 FOR THIS APE 2 FOR THIS REDUCED 414 00:19:00,760 --> 00:19:03,320 ATR DDR PATHWAY IN LSS. 415 00:19:03,320 --> 00:19:04,600 TO FIGURE OUT WHAT'S THE 416 00:19:04,600 --> 00:19:09,160 MECHANISM OF THIS, WE USE 2 417 00:19:09,160 --> 00:19:11,840 TYPES OF MUTATIONS. 418 00:19:11,840 --> 00:19:16,080 ONE IS IN THE PAPER BOX THAT MAY 419 00:19:16,080 --> 00:19:18,280 MEDIATE THE [INDISCERNIBLE] WE 420 00:19:18,280 --> 00:19:21,880 ALSO TESTED TO 2 NUCLEUS 421 00:19:21,880 --> 00:19:25,800 DEFICIENT MUTANT E34 A OR 422 00:19:25,800 --> 00:19:26,520 [INDISCERNIBLE] MUTATION. 423 00:19:26,520 --> 00:19:29,280 SO COMPARED TO THE WILD-TYPE 424 00:19:29,280 --> 00:19:32,760 THIS MUTATION CANNOT RESCUE 425 00:19:32,760 --> 00:19:34,480 CHECK 1 PHOSPHORYLATION BECAUSE 426 00:19:34,480 --> 00:19:36,760 IT CANNOT SUCCESSFULLY RECRUIT 427 00:19:36,760 --> 00:19:39,600 IT FROM THE DNA, THERE'S NO 428 00:19:39,600 --> 00:19:42,240 RECRUITMENT TO ATR COMPLEXES TO 429 00:19:42,240 --> 00:19:45,320 DAMAGE CHROMATIN EITHER, WHEN WE 430 00:19:45,320 --> 00:19:47,200 COMPARE THE NUCLEUS DEFICIENT 431 00:19:47,200 --> 00:19:49,600 MUTATION, WE FOUND WE CANNOT 432 00:19:49,600 --> 00:19:51,280 RESCUE [INDISCERNIBLE], EVEN 433 00:19:51,280 --> 00:19:53,240 THOUGH THE RECRUITMENT TO THE 434 00:19:53,240 --> 00:19:55,680 CHROMATIN DNA HERE IS SAME AS 435 00:19:55,680 --> 00:19:56,960 WILD-TYPE BUT BECAUSE OF THE 436 00:19:56,960 --> 00:20:00,640 DEFICIENCY OF THE NUCLEUS, WE 437 00:20:00,640 --> 00:20:03,720 CANNOT RESCUE THE ATR COMPLEX OF 438 00:20:03,720 --> 00:20:06,400 THE CHROMATIN DNA. 439 00:20:06,400 --> 00:20:09,600 SO THERE ARE 2 MECHANISMS, 1 IS 440 00:20:09,600 --> 00:20:10,960 [INDISCERNIBLE] INTACT FOR THE 441 00:20:10,960 --> 00:20:14,520 BOX, IMPORTANT FOR RECRUITMENT 442 00:20:14,520 --> 00:20:18,040 AND THEN IT'S EXNUCLEIC ACTIVITY 443 00:20:18,040 --> 00:20:19,280 IS IMPORTANT FOR THE DRIVE 444 00:20:19,280 --> 00:20:19,640 DIRECTION. 445 00:20:19,640 --> 00:20:23,600 SO THE MECHANISM FOR THE APE2, 446 00:20:23,600 --> 00:20:25,040 THE ATR, DDR POGHT WAY. 447 00:20:25,040 --> 00:20:27,440 NEXT WITH SCOTT WILLIAMS FROM 448 00:20:27,440 --> 00:20:33,320 NIEHS, VISITED US BACK IN 2000, 449 00:20:33,320 --> 00:20:34,880 2013 OR 14, WE CAME TO 450 00:20:34,880 --> 00:20:39,320 COLLABORATE ON THIS PROJECT AND 451 00:20:39,320 --> 00:20:46,320 WE FOUND THIS VERY UNIQUE MOTIF, 452 00:20:46,320 --> 00:20:49,760 SO MY STUDENT ZACH AND LIN IS 453 00:20:49,760 --> 00:20:52,560 WORKED ON THIS PROJECT BECAUSE 454 00:20:52,560 --> 00:20:56,280 THIS IS ROUGH LOW IN 100 GENES 455 00:20:56,280 --> 00:20:58,640 OR METABOLISM PROTEINS INCLUDING 456 00:20:58,640 --> 00:21:01,560 NIO3, TOP 3 A, SO THEY ARE 457 00:21:01,560 --> 00:21:03,040 HIGHLY CONSERVED IMMUNO ASSAYS. 458 00:21:03,040 --> 00:21:06,480 AND IN THE APE2 IN DIFFERENT 459 00:21:06,480 --> 00:21:08,680 SPECIES, THEY'RE ALSO HIGHLY 460 00:21:08,680 --> 00:21:09,080 CONSERVED. 461 00:21:09,080 --> 00:21:14,680 SO SCOTT IS EXPERT IN STRUCTURE 462 00:21:14,680 --> 00:21:17,640 BIOLOGY SO HIS LAB CRYSTALLIZE 463 00:21:17,640 --> 00:21:21,000 THIS APETO THE FGF MOTIF, IT HAS 464 00:21:21,000 --> 00:21:22,600 UNIQUE HELIOS POSITIVE 465 00:21:22,600 --> 00:21:25,640 EXCITATORY AND ISO HAD 3 BETA 466 00:21:25,640 --> 00:21:28,960 SHADE FORMING THIS WIDE-LIKE 467 00:21:28,960 --> 00:21:31,840 STRUCTURE AND MORE IMPORTANTLY 468 00:21:31,840 --> 00:21:33,840 THIS CAN PREFERENTIALLY BIND TO 469 00:21:33,840 --> 00:21:37,200 SINGLE STRAND DNA AND 1 IMMUNO 470 00:21:37,200 --> 00:21:39,600 ASIGNIFY TO THE MUTATION CAN 471 00:21:39,600 --> 00:21:42,320 COMPROMISE AND PAIR THESE SINGLE 472 00:21:42,320 --> 00:21:43,560 STRAND DNA BINDING. 473 00:21:43,560 --> 00:21:46,920 WHEN WE LOCATED THE X-ACTIVITY 474 00:21:46,920 --> 00:21:57,360 OF THIS USING THIS AS A 475 00:22:06,840 --> 00:22:08,400 SUBSTRATE--OR FILE THE MUTATION, 476 00:22:08,400 --> 00:22:12,080 IT CANNOT BE RESCUED AND COMPLEX 477 00:22:12,080 --> 00:22:15,120 CANNOT BE RESCUED SO THIS 478 00:22:15,120 --> 00:22:18,560 INDICATE THE MOTIF WITHIN THE 479 00:22:18,560 --> 00:22:21,760 AP2 IS CRITICAL FOR IT'S AXO 480 00:22:21,760 --> 00:22:23,400 ACTIVITY AND ALSO IMPORTANT IF 481 00:22:23,400 --> 00:22:27,040 YOU ARE FOR THE ATR DDR 482 00:22:27,040 --> 00:22:27,720 ACTIVATION. 483 00:22:27,720 --> 00:22:32,040 SO THERE ARE 2 QUESTIONS HERE. 484 00:22:32,040 --> 00:22:35,920 WE USE PRIMARILY SO HYDROGEN 485 00:22:35,920 --> 00:22:38,040 PEROXIDE, NOT ONLY IN SINGLE 486 00:22:38,040 --> 00:22:39,480 STRAND BREAKS COULD BE OTHER 487 00:22:39,480 --> 00:22:41,760 TYPES OF THESE LESIONS AS WELL 488 00:22:41,760 --> 00:22:43,160 SO WHAT HAPPENS FOR THE 489 00:22:43,160 --> 00:22:44,760 [INDISCERNIBLE] OF THESE 490 00:22:44,760 --> 00:22:45,880 STRUCTURES? 491 00:22:45,880 --> 00:22:50,640 SO SECOND QUESTION IS HOW IS 492 00:22:50,640 --> 00:22:52,560 THAT SINGLE STRAND GENERATED IF 493 00:22:52,560 --> 00:22:54,760 THIS IS CRITICAL FOR ACTIVATION 494 00:22:54,760 --> 00:22:57,040 OF ATR NUCLEUS ACTIVITY. 495 00:22:57,040 --> 00:23:01,240 TO ADDRESS THE FIRST QUESTION, 496 00:23:01,240 --> 00:23:03,400 BOTH THE POAT DOC IN THE LAB TRY 497 00:23:03,400 --> 00:23:06,200 TO ANSWER THOSE 2 QUESTIONS. 498 00:23:06,200 --> 00:23:09,440 DOES A SINGLE STRAND BREAK 499 00:23:09,440 --> 00:23:10,840 TRIGGER DDR PATH ACTIVATION, 500 00:23:10,840 --> 00:23:12,120 WHAT'S THE MOLECULE OR MECHANISM 501 00:23:12,120 --> 00:23:17,320 OF THIS AS BEING INDUCED FOR THE 502 00:23:17,320 --> 00:23:19,960 DDR PATHWAY. 503 00:23:19,960 --> 00:23:22,120 WE PLASMA BASED USE THIS ENZYME 504 00:23:22,120 --> 00:23:25,720 CAN CREATE SINGLE STRAND BREAK 505 00:23:25,720 --> 00:23:29,400 ON THE TOP STRAND AT THE FINAL 506 00:23:29,400 --> 00:23:32,480 LOCATION WE USE THE PH OS, OF 507 00:23:32,480 --> 00:23:36,880 THE PHOSPHATE SO BOTH ARE OH ON 508 00:23:36,880 --> 00:23:37,400 THE 3 AND 5 SIDE. 509 00:23:37,400 --> 00:23:39,200 SO THIS IS CALLED CONTROL PLAZ 510 00:23:39,200 --> 00:23:41,520 MIDS MID, THIS IS SSB PLAZ MIDS 511 00:23:41,520 --> 00:23:44,520 MID, SO IF YOU USE THE ENZYME TO 512 00:23:44,520 --> 00:23:50,080 LINEARIZE THIS, WE CALL THIS 513 00:23:50,080 --> 00:23:52,800 STRUCTURES BECAUSE THIS ENZYME 514 00:23:52,800 --> 00:23:54,120 GENERATE NYK CANNOT BE 515 00:23:54,120 --> 00:23:58,000 RECOGNIZED BY THE SBF ENZYME 516 00:23:58,000 --> 00:23:58,240 ANIMAL. 517 00:23:58,240 --> 00:24:00,320 SO THIS IS EXPECTED. 518 00:24:00,320 --> 00:24:02,440 USING THE SSB PLAZ MIDS MID, 519 00:24:02,440 --> 00:24:04,520 COMPARE THIS CONTROL PLAZ MIDS 520 00:24:04,520 --> 00:24:08,040 MID, WE ADD DIFFERENT DOSES FOR 521 00:24:08,040 --> 00:24:10,480 THESE S, B COME INTO HS WE SEE. 522 00:24:10,480 --> 00:24:14,560 WE SEE DOSE DEPENDENT ACTIVATION 523 00:24:14,560 --> 00:24:17,280 OF CHK 1 ACTIVATION, WE FURGTDER 524 00:24:17,280 --> 00:24:19,640 VALIDATE USING AT TR INHIBITOR, 525 00:24:19,640 --> 00:24:30,160 822 THAT IS COMPROMISED BUT AND 526 00:24:40,360 --> 00:24:41,520 DEPLETED THOSE TECH POINT 527 00:24:41,520 --> 00:24:44,960 PROTEIN, WE FOUND ASB INDUCED 528 00:24:44,960 --> 00:24:46,640 CHECK POINT PHOSPHORYLATION ALL 529 00:24:46,640 --> 00:24:51,640 COMPROMISED, SO WE ARE PRETTY 530 00:24:51,640 --> 00:24:56,440 SURE THIS IS AND THE SSB 531 00:24:56,440 --> 00:25:03,600 PLASMID, FURTHER MORE WE 532 00:25:03,600 --> 00:25:07,240 CHARACTER AND MODEL THE MOTIF, 533 00:25:07,240 --> 00:25:09,960 WITH APE PIVOT BOX, THIS NEW 534 00:25:09,960 --> 00:25:12,360 MODE, WE CALL MODE 2 THAT 535 00:25:12,360 --> 00:25:16,720 MEDIATED BETWEEN THE APE-TO THE 536 00:25:16,720 --> 00:25:19,640 MOTIF TO THE AND THE PCA AND 537 00:25:19,640 --> 00:25:23,560 THIS MODEL 2 INTAKS IS CRITICAL 538 00:25:23,560 --> 00:25:34,040 BECAUSE C-4708 WILL LOSE THE 539 00:26:01,280 --> 00:26:03,560 INTAKS--REPAIR BACK TO THE 540 00:26:03,560 --> 00:26:06,720 DOUBLE STRANDA PLASMID DNA 541 00:26:06,720 --> 00:26:12,600 TAKEN--THEY AREY SOUV THE 542 00:26:12,600 --> 00:26:13,920 SENSITIVITY, AND THESE ARE THE 543 00:26:13,920 --> 00:26:20,040 SSB PAIR THAT CAN BE QUANTIFIED, 544 00:26:20,040 --> 00:26:24,120 AND WHEN THAT IS TREATED PERFECT 545 00:26:24,120 --> 00:26:27,640 WITH ATR INHIBITED, THOSE REPAIR 546 00:26:27,640 --> 00:26:30,200 PRODUCTS WILL BE COMPROMISED, 547 00:26:30,200 --> 00:26:33,320 SIMILARLY, WHEN WE DEPLETE APE2, 548 00:26:33,320 --> 00:26:36,640 CHECK 1, ATR CHECK 1 CANNOT BE 549 00:26:36,640 --> 00:26:39,840 ACTIVATED SO THE SSB REPAIR IS 550 00:26:39,840 --> 00:26:41,520 COMPROMISED, WE ADD BACK THE 551 00:26:41,520 --> 00:26:43,840 MAKE APE TO PROTEIN, THEN THE 552 00:26:43,840 --> 00:26:45,920 SSB REPAIR MAY BE RESUMED, SO 553 00:26:45,920 --> 00:26:49,800 THIS IS A QUANTIFICATION FACT, 554 00:26:49,800 --> 00:26:52,400 SO, BOTH ATR IS APE2 ARE 555 00:26:52,400 --> 00:26:53,720 ESSENTIAL FOR SSB REPAIR. 556 00:26:53,720 --> 00:26:59,280 NOW HERE ARE THE MECHANISMS OR 557 00:26:59,280 --> 00:27:04,920 THE SSB PAIR MEDIATED BY APE2. 558 00:27:04,920 --> 00:27:10,640 SO THE SSB CAN RECRUIT APE2, 559 00:27:10,640 --> 00:27:13,480 AFTER RECRUITMENT APECAN BE 560 00:27:13,480 --> 00:27:15,360 ACTIVATED AND ONCE APECHOOSE 561 00:27:15,360 --> 00:27:18,720 ACTIVATED IT CAN PERFORM 562 00:27:18,720 --> 00:27:19,920 ACTIVITY THROUGH 5 DIRECTION, 563 00:27:19,920 --> 00:27:23,160 THROUGH SINGLE STRAND DNA, CODED 564 00:27:23,160 --> 00:27:25,320 WITH COMPLEX, IT WILL BE 565 00:27:25,320 --> 00:27:27,280 RECORDED, THEN PROMOTE SSB TO BE 566 00:27:27,280 --> 00:27:27,760 REPAIRED. 567 00:27:27,760 --> 00:27:29,840 SO SUMMARIZE SO FAR WE HAVE 568 00:27:29,840 --> 00:27:32,600 THERE ARE 3 MECHANISM FOR APE2 569 00:27:32,600 --> 00:27:34,360 FUNCTION HERE. 570 00:27:34,360 --> 00:27:38,120 THE FIRST 1 IS MEDIATED BY APE2, 571 00:27:38,120 --> 00:27:40,760 PAPER BOX INTAKS OF THE 572 00:27:40,760 --> 00:27:41,360 [INDISCERNIBLE] WHICH IS 573 00:27:41,360 --> 00:27:43,640 IMPORTANT FOR THE RECRUITMENT OF 574 00:27:43,640 --> 00:27:47,240 APE2 TO THE DAMAGE SIDE, SECOND 575 00:27:47,240 --> 00:27:50,160 MECHANISM IS APE20, FOR THE 576 00:27:50,160 --> 00:27:51,800 MOTIF, CAN INTACT WITH THE 577 00:27:51,800 --> 00:27:52,960 SINGLE STRAND DNA THAT'S 578 00:27:52,960 --> 00:27:55,360 IMPORTANT FOR ACTIVITY IN THE 579 00:27:55,360 --> 00:27:55,840 ACTIVITY. 580 00:27:55,840 --> 00:28:00,560 THIRD MECHANISM IS, THE APE, TO 581 00:28:00,560 --> 00:28:03,080 EGRF MOTIF INTACT WITH THE 582 00:28:03,080 --> 00:28:05,680 [INDISCERNIBLE] POETIC TEEF, 583 00:28:05,680 --> 00:28:09,560 THAT'S ALSO IMPORTANT FOR ATI 584 00:28:09,560 --> 00:28:10,040 VACTIVATION. 585 00:28:10,040 --> 00:28:11,200 SO THERE ARE NEXT SEVERAL 586 00:28:11,200 --> 00:28:12,280 QUESTIONS RESOLVED HERE SO WE 587 00:28:12,280 --> 00:28:17,160 TRY TO DO MORE BY ANALYSIS TO 588 00:28:17,160 --> 00:28:18,680 ATP TO EXPRESSION IN CANCER AND 589 00:28:18,680 --> 00:28:21,120 WE ALSO WANT TO KNOW HOW DOES 590 00:28:21,120 --> 00:28:23,960 APE CONTRIBUTE TO CANCER 591 00:28:23,960 --> 00:28:24,240 ETIOLOGY? 592 00:28:24,240 --> 00:28:27,920 IS APE A DRIVER OR PASSENGER OF 593 00:28:27,920 --> 00:28:28,120 CANCER? 594 00:28:28,120 --> 00:28:30,720 WE STILL HAVE A LOT OF THINGS TO 595 00:28:30,720 --> 00:28:33,040 DO WITH THIS, THE QUESTION IS 596 00:28:33,040 --> 00:28:35,920 CAN APE BE TARGETED FOR CANCER 597 00:28:35,920 --> 00:28:36,160 TREATMENT. 598 00:28:36,160 --> 00:28:38,040 WE WILL TELL YOU SOME PROGRESS 599 00:28:38,040 --> 00:28:42,040 ON THOSE DIRECTIONS. 600 00:28:42,040 --> 00:28:47,840 IN LITERATURE, APE OR EXPRESSION 601 00:28:47,840 --> 00:28:49,160 HAS BEEN [INDISCERNIBLE] 602 00:28:49,160 --> 00:28:56,360 MULTIMELANOMA MN, THIS IS AP TO 603 00:28:56,360 --> 00:28:57,480 MRN EXPRESSION, IT'S OVER 604 00:28:57,480 --> 00:29:01,840 EXPRESSED IN PATIENTS IN THE 605 00:29:01,840 --> 00:29:04,240 MM LEVEL THIS ALSO SHOWED APE 606 00:29:04,240 --> 00:29:07,760 EXPRESSION OR APE 2. 607 00:29:07,760 --> 00:29:09,680 ANOTHER STUDY SHOWS IN LABOR 608 00:29:09,680 --> 00:29:10,760 CANCER PATIENT, THE LEVEL 609 00:29:10,760 --> 00:29:11,760 EXPRESSION, THE TUMOR COMPARED 610 00:29:11,760 --> 00:29:17,600 IT WITH A NORMAL TISSUE SYSTEM 611 00:29:17,600 --> 00:29:18,760 ALSO OVEREXPRESSED. 612 00:29:18,760 --> 00:29:21,400 MORE IMPORTANTLY, APE TO 613 00:29:21,400 --> 00:29:24,760 OVEREXPRESS OR HIGHLY APE 2 614 00:29:24,760 --> 00:29:27,080 EXPRESSION ASSOCIATED WITH THIS 615 00:29:27,080 --> 00:29:34,200 LOWER OVERALL SURVIVAL RATE IN 616 00:29:34,200 --> 00:29:36,080 CANCER PATIENTS. 617 00:29:36,080 --> 00:29:39,120 SO COLLABORATING WITH MY 618 00:29:39,120 --> 00:29:42,160 COLLEAGUE AT UNC SHE MOVED TO 619 00:29:42,160 --> 00:29:44,920 [INDISCERNIBLE] AND AT 620 00:29:44,920 --> 00:29:45,520 UNIVERSITY AND [INDISCERNIBLE] 621 00:29:45,520 --> 00:29:46,680 WORKED ON THIS DRUG AND FOUND 622 00:29:46,680 --> 00:29:49,760 OUT THAT FROM THE TCGA ANALYSIS, 623 00:29:49,760 --> 00:29:55,120 WE FOUND IN 14 CANCER TYPES 624 00:29:55,120 --> 00:29:57,240 ROUGHLY 17% FREQUENCY OF THE 625 00:29:57,240 --> 00:30:00,760 NORMAL ALTERATIONS, ALSO THOSE 626 00:30:00,760 --> 00:30:05,200 OLTERATIONS INCLUDING GAMES OR 627 00:30:05,200 --> 00:30:07,280 AMPLIFICATION, OR THE HOMOLOGY 628 00:30:07,280 --> 00:30:09,880 DIVISION OF APE2 GENE FROM THOSE 629 00:30:09,880 --> 00:30:11,920 CANCER PATIENTS. 630 00:30:11,920 --> 00:30:16,240 ROUGHLY 120 SOMATIC MUTATIONS IN 631 00:30:16,240 --> 00:30:18,720 2 DIFFERENT CANCER TYPES ACROSS 632 00:30:18,720 --> 00:30:20,840 THE WHOLE CODING GENE, SO APE2 633 00:30:20,840 --> 00:30:24,000 IN DIFFERENT REGIONS INCLUDING 634 00:30:24,000 --> 00:30:27,880 THE [INDISCERNIBLE] ZF GF MOTIF. 635 00:30:27,880 --> 00:30:30,200 WHEN WE COMPARED THE MATCHED 636 00:30:30,200 --> 00:30:34,680 TISSUE, TUMOR TISSUE VERSUS THE 637 00:30:34,680 --> 00:30:36,480 NONMALIGNANT TISSUES WE FOUND 6 638 00:30:36,480 --> 00:30:40,280 CANCER TYPES HIDDEN IN THE 639 00:30:40,280 --> 00:30:41,640 LABOR, [INDISCERNIBLE] ALONG 640 00:30:41,640 --> 00:30:46,160 APE2 IS OVEREXPRESSED IN TUMOR 641 00:30:46,160 --> 00:30:46,960 TISSUES. 642 00:30:46,960 --> 00:30:49,800 AND THE EXPRESSION ALSO 643 00:30:49,800 --> 00:30:52,000 CORRELATES WELL WITH OTHER 644 00:30:52,000 --> 00:30:54,720 IMPAIRED DDR PATHWAYS INCLUDING 645 00:30:54,720 --> 00:30:58,440 [INDISCERNIBLE] P A RP AND 646 00:30:58,440 --> 00:30:59,600 OTHERS. 647 00:30:59,600 --> 00:31:02,240 SO WHEN WE USE PANCREATIC CANCER 648 00:31:02,240 --> 00:31:05,760 CELLS WE LOOK AT THE DIFFERENT 649 00:31:05,760 --> 00:31:08,520 ATR REQUIRING ACTIVATION BY 650 00:31:08,520 --> 00:31:13,840 DIFFERENT STRESSORS INCLUDING 651 00:31:13,840 --> 00:31:16,000 HYDROGEN PEROXIDE, 652 00:31:16,000 --> 00:31:16,480 [INDISCERNIBLE], 653 00:31:16,480 --> 00:31:18,760 [INDISCERNIBLE], ETO, AND THE 654 00:31:18,760 --> 00:31:20,200 INHIBITOR FOR THE 655 00:31:20,200 --> 00:31:20,560 [INDISCERNIBLE]. 656 00:31:20,560 --> 00:31:22,480 THE CHECK 1 PHOSPHORYLATION BY 657 00:31:22,480 --> 00:31:27,560 THE STRESSFUL CONDITIONS, AND IN 658 00:31:27,560 --> 00:31:30,480 THE RP2 STUDY ARE COMPROMISED 659 00:31:30,480 --> 00:31:32,640 WITH THE APEPROTEIN IS KNOCKED 660 00:31:32,640 --> 00:31:35,680 DOWN BY SiRNA MEDIATED 661 00:31:35,680 --> 00:31:37,400 APCEREBELLUMS 2 KNOCK DOWN. 662 00:31:37,400 --> 00:31:40,040 WE ALSO QUANTIFIED THE GAMMA TO 663 00:31:40,040 --> 00:31:46,920 THE X, WE FOUND APETO KNOCK 664 00:31:46,920 --> 00:31:48,080 DOWN, THE ELEVATED, AND THEY'RE 665 00:31:48,080 --> 00:31:50,400 UNTREATED OR THE STRESSFUL 666 00:31:50,400 --> 00:31:51,760 CONDITIONS AND ALSO 667 00:31:51,760 --> 00:31:54,960 [INDISCERNIBLE] APE TO SENSITIZE 668 00:31:54,960 --> 00:31:57,200 CANCER CELL TO THOSE 4 DIFFERENT 669 00:31:57,200 --> 00:32:01,520 TYPES OF DRUGS OR STRESSFUL 670 00:32:01,520 --> 00:32:01,800 CONDITIONS. 671 00:32:01,800 --> 00:32:06,280 AND TO MOVE THIS FORWARD, WE ARE 672 00:32:06,280 --> 00:32:07,800 INTERESTED IN THE APPLICATION OF 673 00:32:07,800 --> 00:32:10,080 SPECIFIC INPIB HIB THORSITTOR 674 00:32:10,080 --> 00:32:14,480 FROM 1 PAPER PUBLISHED IN 2014, 675 00:32:14,480 --> 00:32:16,240 THERE'S SEVERAL DRUGS, 676 00:32:16,240 --> 00:32:19,320 IDENTIFIED TO INHIBIT ATR DDR 677 00:32:19,320 --> 00:32:21,880 PATHWAY, SO WE MADE HYPOTHESIS, 678 00:32:21,880 --> 00:32:25,200 WHILE IT'S CLOSE TO STRUCTURALLY 679 00:32:25,200 --> 00:32:31,360 THE [INDISCERNIBLE], WE TEST 680 00:32:31,360 --> 00:32:32,600 WHETHER CELASTROL CAN 681 00:32:32,600 --> 00:32:38,560 IBT--INTEGRATE HIB THORSIT THIS 682 00:32:38,560 --> 00:32:43,360 SSB INDUCED SYSTEM, WE ALSO 683 00:32:43,360 --> 00:32:45,520 FOUND THIS CELASTROL CAN INHIBIT 684 00:32:45,520 --> 00:32:48,320 THE MOTIF COMPLEX OVER THE 685 00:32:48,320 --> 00:32:52,120 SINGLE STRAND DNA, ALSO 686 00:32:52,120 --> 00:32:55,240 CELASTROL INHIBITS THE, DOSE 687 00:32:55,240 --> 00:32:57,080 DEPENDENTS THE ACTIVITY. 688 00:32:57,080 --> 00:33:00,560 SO WE IDENTIFY CELASTROL IS THE 689 00:33:00,560 --> 00:33:03,400 FIRST KNOWN COMPOUND INHIBITOR 690 00:33:03,400 --> 00:33:07,800 OF APE, 2, WE ALSO TEST WHETHER 691 00:33:07,800 --> 00:33:09,200 THE CELASTROL TREATMENT CAN 692 00:33:09,200 --> 00:33:10,600 SYNTHESIZE TREATMENT OR CANCER 693 00:33:10,600 --> 00:33:13,560 CELLS TO THOSE STRESSFUL 694 00:33:13,560 --> 00:33:15,720 CONDITIONS, TO INCLUDING 695 00:33:15,720 --> 00:33:20,120 KD--SALLY WHARKS CPT AND THE 696 00:33:20,120 --> 00:33:20,480 ETO. 697 00:33:20,480 --> 00:33:26,520 FURTHER MORE I COLLABORATE WITH 698 00:33:26,520 --> 00:33:27,520 [INDISCERNIBLE] FROM THE 699 00:33:27,520 --> 00:33:29,680 CLEVELAND CLINIC TO LOOK AT APE 700 00:33:29,680 --> 00:33:31,960 TO FUNCTION IN MOUSE AND THE 701 00:33:31,960 --> 00:33:33,040 MITOCHONDRIA STABILITY. 702 00:33:33,040 --> 00:33:36,840 AS YOU KNOW, SIS PLATIN CAN 703 00:33:36,840 --> 00:33:38,800 CAUSE ACUTE INJURY CALLED AKI, 704 00:33:38,800 --> 00:33:44,000 IT'S A SIDE EFFECTS OF THE 705 00:33:44,000 --> 00:33:46,040 TREATMENT, IT WAS IMPORTANT WE 706 00:33:46,040 --> 00:33:49,960 FOUND INDUCED AKI, IF MEDIATED 707 00:33:49,960 --> 00:33:50,600 FROM APE2 UPREGULATION. 708 00:33:50,600 --> 00:33:55,720 SO THIS IS A DIAGRAM TO SHOW THE 709 00:33:55,720 --> 00:33:58,880 PROBLEM CISPLATIN HAPPENS AT A 710 00:33:58,880 --> 00:34:00,720 UNIQUE [INDISCERNIBLE] CALLED 711 00:34:00,720 --> 00:34:01,960 PROXIMAL TUBAL CELL IN 712 00:34:01,960 --> 00:34:06,480 [INDISCERNIBLE], IN THOSE CELLS 713 00:34:06,480 --> 00:34:09,800 SIT CISPLATEIN TREATMENT CAN 714 00:34:09,800 --> 00:34:11,480 CAUSE APE2 UPREGULATION AND CAN 715 00:34:11,480 --> 00:34:15,280 CAUSE CROSS LINK DAMAGE IN DNA 716 00:34:15,280 --> 00:34:16,600 WHERE INTERESTINGLY THE 717 00:34:16,600 --> 00:34:19,760 UPREGULATED APE CAN GET INTO THE 718 00:34:19,760 --> 00:34:21,480 MT AND INTACT WITH KNOWN MUSCLE 719 00:34:21,480 --> 00:34:25,160 MOUSE IN PROTEIN CALLED M1 A9. 720 00:34:25,160 --> 00:34:29,120 THAT LIKELY IS THE REASON FOR 721 00:34:29,120 --> 00:34:32,800 CISPLATIN INDUCED AKI. 722 00:34:32,800 --> 00:34:35,240 THIS IS NORMAL PATIENTS VERSUS 723 00:34:35,240 --> 00:34:38,720 AFTER THE TREATMENT OR THE CIS 724 00:34:38,720 --> 00:34:41,320 PLATIN API, THE IT'S 725 00:34:41,320 --> 00:34:43,000 UNREGULATED, OVER HERE IS 726 00:34:43,000 --> 00:34:46,600 CISPLATIN TREATMENT IN THE MOUSE 727 00:34:46,600 --> 00:34:49,320 THAT TREATMENT CAN CAUSE APE TO 728 00:34:49,320 --> 00:34:51,320 UPREGULATION FOR QUANTIFICATION 729 00:34:51,320 --> 00:34:55,880 BUT NOT APE1 UPREGULATION. 730 00:34:55,880 --> 00:34:57,840 THE [INDISCERNIBLE] GROUP HAVE 731 00:34:57,840 --> 00:35:01,360 THIS KIND OF APE EXPRESSION, 732 00:35:01,360 --> 00:35:04,160 TRANSGENIC MICE SO THIS IS THE 733 00:35:04,160 --> 00:35:08,440 NORMAL CONTROL, THIS IS APE 734 00:35:08,440 --> 00:35:10,240 OVERXRETIONZ, GLOBAL EXPRESSION 735 00:35:10,240 --> 00:35:13,320 THAT CAN CAUSE MORE EARLY DEATHS 736 00:35:13,320 --> 00:35:18,680 OF THE MOUSE, AND WHEN WIEW LOOK 737 00:35:18,680 --> 00:35:20,600 AT THE MOUSE ANALYSIS, COMPARE 738 00:35:20,600 --> 00:35:26,240 WITH THE CONTROL VERSUS APE TO 739 00:35:26,240 --> 00:35:27,840 OVEREXPRESSION, THEY FOUND THE 740 00:35:27,840 --> 00:35:29,560 EXPRESSION CAN CAUSE MORE 741 00:35:29,560 --> 00:35:31,400 CASPACE 3 AND GAMMA 742 00:35:31,400 --> 00:35:34,040 [INDISCERNIBLE] INDICATE MORE 743 00:35:34,040 --> 00:35:37,360 DNA DAMAGE OR CELL DEATH. 744 00:35:37,360 --> 00:35:39,120 WE LOOK AT THE MILD DON DID 745 00:35:39,120 --> 00:35:42,640 YOUIT STRUCTURE WE FOUND THIS 746 00:35:42,640 --> 00:35:45,160 NORMAL MICE, NORMAL MITOCHONDRIA 747 00:35:45,160 --> 00:35:50,120 MORPHOLOGY, FOLLOWING THE APE 748 00:35:50,120 --> 00:35:54,000 OVEREXPRESSED BECOMES ABNORMAL. 749 00:35:54,000 --> 00:35:55,400 CISPLATIN CAUSED [INDISCERNIBLE] 750 00:35:55,400 --> 00:35:58,040 OR OVEREXPRESSION CAUSED MORE 751 00:35:58,040 --> 00:36:01,320 LOCALIZATION OF APETO BE COX4, 752 00:36:01,320 --> 00:36:03,720 OF A MITOCHONDRIA EXPRESSIVE 753 00:36:03,720 --> 00:36:05,200 MARKER AND THE EXPRESSION ALSO 754 00:36:05,200 --> 00:36:15,600 CAN CAUSE MORE ENRICHED 755 00:36:15,960 --> 00:36:16,200 RICHED 756 00:36:16,200 --> 00:36:16,560 COLOCALIZATION. 757 00:36:16,560 --> 00:36:18,240 SO THIS IS A SUMMARY HERE, THIS 758 00:36:18,240 --> 00:36:22,400 IS THE MOTIF THAT ARE REQUIRED 759 00:36:22,400 --> 00:36:24,560 FOR EXPRESS-DUCED ATR, DDR 760 00:36:24,560 --> 00:36:26,480 PATHWAY AND APE 2 TOAXON 761 00:36:26,480 --> 00:36:29,120 NUCLEACE ACTIVITY IS REQUIRED 762 00:36:29,120 --> 00:36:31,000 FOR SSB INDUCED ATR AND DDR 763 00:36:31,000 --> 00:36:33,600 PATHWAY AND THE SSB PAIRS, THE 764 00:36:33,600 --> 00:36:35,560 FUNCTIONAL APE TO ATR, DID, DR 765 00:36:35,560 --> 00:36:42,360 AND I CAN TARGET FOR CANCER 766 00:36:42,360 --> 00:36:42,920 TREATMENT. 767 00:36:42,920 --> 00:36:49,160 SO WE PROPOSE APETO SIGNIFICANT 768 00:36:49,160 --> 00:36:51,040 FOR THAT, FOR--ESPECIALLY IN 769 00:36:51,040 --> 00:36:52,960 BRCA 1 AND 2 DEFICIENT CELLS. 770 00:36:52,960 --> 00:36:56,280 THAT IS A SUMMARY OF OUR 771 00:36:56,280 --> 00:36:57,640 PUBLICATIONS OF APE OVER THE 772 00:36:57,640 --> 00:36:58,040 YEARS. 773 00:36:58,040 --> 00:37:01,200 NEXT I WILL SWITCH GEAR TO APE1, 774 00:37:01,200 --> 00:37:04,040 THE OLDER BROTHER, HOW APE 775 00:37:04,040 --> 00:37:07,040 FUNCTION AS SSB INDUCED ATR, DDR 776 00:37:07,040 --> 00:37:11,160 AND HOW THAT MEDIATE ATR 777 00:37:11,160 --> 00:37:11,680 SIGNALING AND UNEXPECTED 778 00:37:11,680 --> 00:37:13,840 FUNCTIONING IN THE ALL THE DDR 779 00:37:13,840 --> 00:37:14,200 ACTIVATION. 780 00:37:14,200 --> 00:37:23,320 SO I SHARE WITH YOU, APE IS 781 00:37:23,320 --> 00:37:27,160 RECRUITED AND ACTIVATED AT 782 00:37:27,160 --> 00:37:29,960 SSB SITE, FOR THE TUITION, WE 783 00:37:29,960 --> 00:37:32,320 HYPOTH SIZE THAT APE 2 AND APE 1 784 00:37:32,320 --> 00:37:35,680 PLAYED A DIRECT ROLE IN THIS 785 00:37:35,680 --> 00:37:39,640 SIGNALING BECAUSE APE HAS SLOW 786 00:37:39,640 --> 00:37:42,720 OR WEAK ACTIVITY GENERATED THAT 787 00:37:42,720 --> 00:37:45,720 SINGLE STRAND WILL GET. 788 00:37:45,720 --> 00:37:50,960 FIRST OF ALL, THIS STUDY WAS 789 00:37:50,960 --> 00:37:52,120 CONDUCTED BY [INDISCERNIBLE], 790 00:37:52,120 --> 00:37:55,720 AND AN HONORS STUDENT JUDE ON 791 00:37:55,720 --> 00:37:58,120 THIS PROJECT, SO THE FIRST 792 00:37:58,120 --> 00:37:59,960 QUESTION WILL TEST WHETHER WE 793 00:37:59,960 --> 00:38:03,480 DEPLETE APE 1 DOES THAT OOSKT 794 00:38:03,480 --> 00:38:05,840 APE TO RECORD TO THE SITE? 795 00:38:05,840 --> 00:38:09,200 SO WE USE THIS APE TO PREDICT 796 00:38:09,200 --> 00:38:12,440 THE ANTIBODY APE1, SO NO APE 797 00:38:12,440 --> 00:38:15,520 WILL BE REPORTED, TO THE SSB 798 00:38:15,520 --> 00:38:17,760 SITE, THEN APE2 CANNOT BE 799 00:38:17,760 --> 00:38:18,280 RECORDED. 800 00:38:18,280 --> 00:38:21,200 HOWEVER, IF WE DEPLETE APE2, 801 00:38:21,200 --> 00:38:23,920 THEN APE1 STILL CAN BE RECRUITED 802 00:38:23,920 --> 00:38:27,760 TO SSB SITE, INDICATING THAT APE 803 00:38:27,760 --> 00:38:29,720 1 IS UPSTREAM OR APE TO 804 00:38:29,720 --> 00:38:30,160 RECRUITMENT. 805 00:38:30,160 --> 00:38:32,240 SO THIS IS QUITE INTERESTING 806 00:38:32,240 --> 00:38:35,400 BECAUSE P A RP 1 HAS BEEN ISSUED 807 00:38:35,400 --> 00:38:37,280 AS THE SENSOR OF [INDISCERNIBLE] 808 00:38:37,280 --> 00:38:42,560 SO WE USE THIS M-SIZE TO COMPARE 809 00:38:42,560 --> 00:38:44,960 DIFFERENT SUBSTRATES BINDING BY 810 00:38:44,960 --> 00:38:46,120 APE 1. 811 00:38:46,120 --> 00:38:48,320 WE USE DOUBLE STRAND DNA WITH A 812 00:38:48,320 --> 00:38:50,720 SINGLE STRAND BREAK OR A SMALL 813 00:38:50,720 --> 00:38:54,440 [INDISCERNIBLE] SO WE SEE THIS 814 00:38:54,440 --> 00:38:57,080 KIND OF DOSE DEPENDENT ASSEMBLY 815 00:38:57,080 --> 00:39:03,120 OF THE SINGLE STRAND DNA WITH 816 00:39:03,120 --> 00:39:05,440 THIS APE 1 P A RP WHERE THE 817 00:39:05,440 --> 00:39:09,760 SHIFT AFTER BINDING TO SINGLE 818 00:39:09,760 --> 00:39:11,400 STRAIN GAP. 819 00:39:11,400 --> 00:39:13,120 INTERESTINGLY WITH THE GAP 820 00:39:13,120 --> 00:39:15,200 BECOMES 1-3 WITH THE SIDE THEN 821 00:39:15,200 --> 00:39:16,840 THERE'S LESS [INDISCERNIBLE] IF 822 00:39:16,840 --> 00:39:19,920 APE1 AND QUANTIFIED BY MST AS 823 00:39:19,920 --> 00:39:24,520 ALL THIS APE1 BINDING AS A 824 00:39:24,520 --> 00:39:24,840 RESULT. 825 00:39:24,840 --> 00:39:27,840 VERY IMPORTANT TO TRY TO KNOW 826 00:39:27,840 --> 00:39:29,160 WHETHER APE1 CONNECTIVITY IS 827 00:39:29,160 --> 00:39:32,240 ESSENTIAL FOR THIS SIGNALING 828 00:39:32,240 --> 00:39:32,520 PATHWAY. 829 00:39:32,520 --> 00:39:36,360 SO WE IDENTIFY CRITICAL RESIDUE 830 00:39:36,360 --> 00:39:39,360 MUTATED CALLED D306 A WHICH IS 831 00:39:39,360 --> 00:39:45,840 HIGHLY CONSERVED IN THE XEN 832 00:39:45,840 --> 00:39:48,720 OPUS MOUSE, SO THE WILD-TYPE 833 00:39:48,720 --> 00:39:53,160 MAKE APE1 BUT NOT THIS 834 00:39:53,160 --> 00:39:56,000 EXODEFICIENT AND THE COMPLEX 835 00:39:56,000 --> 00:39:57,640 RECRUITMENT TO S'S BESIDES. 836 00:39:57,640 --> 00:40:01,640 AND FOR THE S'S TO BE REPAIRED, 837 00:40:01,640 --> 00:40:04,680 IF YOU DEPLETE IT HAS TO BE 838 00:40:04,680 --> 00:40:05,520 REPAIRED, COMPROMISED BUT WHEN 839 00:40:05,520 --> 00:40:08,280 YOU ADD BACK THE WILD-TYPE AND 840 00:40:08,280 --> 00:40:10,280 NOT THE D MUTANT CAN RESCUE THE 841 00:40:10,280 --> 00:40:11,640 SSB PAIR. 842 00:40:11,640 --> 00:40:22,080 SO APE1 IS FOR THE FOR THE 843 00:40:37,720 --> 00:40:39,080 SIGNALING AS WELL. 844 00:40:39,080 --> 00:40:41,600 INITIATED THE INTERSECTION FOR 845 00:40:41,600 --> 00:40:45,800 THIS, TO GENERATE THE STEP 1. 846 00:40:45,800 --> 00:40:46,480 INITIATION OR INITIATION. 847 00:40:46,480 --> 00:40:53,200 THE SECOND STEP IS APE2 IS 848 00:40:53,200 --> 00:40:56,800 RECRUITED AND ACTIVATED BY THOSE 849 00:40:56,800 --> 00:40:59,360 3 EARLIER TO CONTINUE THE 850 00:40:59,360 --> 00:41:02,120 INTERSECTION SO YOU CAN GET A 851 00:41:02,120 --> 00:41:04,000 LONGER SINGLE STRAND DNA THAN 852 00:41:04,000 --> 00:41:07,640 ATR COMPLEXES WILL BE RECRUITED 853 00:41:07,640 --> 00:41:09,600 THAT YOU EVENTUALLY FILL THE GAP 854 00:41:09,600 --> 00:41:12,520 OF THE SSB REPAIRS COMLICATE. 855 00:41:12,520 --> 00:41:15,200 SO THIS MECHANISM IS ALSO 856 00:41:15,200 --> 00:41:17,120 RECOGNIZED BY COLLEAGUE, KEITH 857 00:41:17,120 --> 00:41:19,320 IN HIS REVIEW, IN THE CELL 858 00:41:19,320 --> 00:41:21,600 BIOLOGY, TIED TO OTHER SINGLE 859 00:41:21,600 --> 00:41:23,720 STRAND BREAK PAIR FOR THE 860 00:41:23,720 --> 00:41:25,760 [INDISCERNIBLE] DISEASES 861 00:41:25,760 --> 00:41:27,400 PUBLISHED LAST YEAR IN 2022. 862 00:41:27,400 --> 00:41:31,120 OKAY, I WANT TO SHARE ANOTHER 863 00:41:31,120 --> 00:41:33,080 UPDATE, VERY, VERY RECENT UPDATE 864 00:41:33,080 --> 00:41:38,480 OF ATR DDR DR SIGNALING PATHWA. 865 00:41:38,480 --> 00:41:40,880 AS YOU ALL LEARNED AROUND 20 866 00:41:40,880 --> 00:41:45,320 YEARS AGO, THIS IS A LEGENDARY 867 00:41:45,320 --> 00:41:47,320 COMPLEX RECRUITED TO DAMAGE SITE 868 00:41:47,320 --> 00:41:49,040 AND MEDIATED ATR ACTIVATION 869 00:41:49,040 --> 00:41:55,920 BREFLY, EITHER OF THE DSB ENDS 870 00:41:55,920 --> 00:41:58,920 OR [INDISCERNIBLE] SO SINGLE 871 00:41:58,920 --> 00:42:02,800 STRAND DNA WILL BE RECRUITED AND 872 00:42:02,800 --> 00:42:06,240 REPORTED AFTER THE SINGLE STRAND 873 00:42:06,240 --> 00:42:09,320 DNA BECAUSE THE RPA-ATRIP 874 00:42:09,320 --> 00:42:14,560 INTERACTION CAN BE RECRUITED TO 875 00:42:14,560 --> 00:42:16,160 THOSE RPA ENIN THE SITES OR IT 876 00:42:16,160 --> 00:42:21,960 IS [INDISCERNIBLE] SO THIS IS 877 00:42:21,960 --> 00:42:24,560 THE TRADITIONAL LEGENDARY MODEL 878 00:42:24,560 --> 00:42:28,600 PROPOSED BY ZOU AND ELLEDGE 20 879 00:42:28,600 --> 00:42:29,520 YEARS AGO. 880 00:42:29,520 --> 00:42:30,760 BUT INTERESTINGLY FOLLOW UP 881 00:42:30,760 --> 00:42:32,920 STUDY FROM OTHER LABS, INCLUDING 882 00:42:32,920 --> 00:42:37,640 [INDISCERNIBLE] AND OTHERS SHOW, 883 00:42:37,640 --> 00:42:39,840 NOT ONLY RPE DEPENDENT BUT ALSO 884 00:42:39,840 --> 00:42:42,640 THE ATR TRIP RECRUITMENT DAMAGE 885 00:42:42,640 --> 00:42:45,720 SITE COULD ALSO BE RPE 886 00:42:45,720 --> 00:42:46,160 INDEPENDENT. 887 00:42:46,160 --> 00:42:49,840 SO WHAT'S THE PROTEIN THAT CAN 888 00:42:49,840 --> 00:42:53,280 BRING ATR COMPLEX TO ASSESS DNA 889 00:42:53,280 --> 00:42:56,720 IN THE RPA INDEPENDENT MANOR FOR 890 00:42:56,720 --> 00:42:58,680 ACTIVATION, HAS BEEN AN 891 00:42:58,680 --> 00:43:00,240 OUTSTANDING QUESTION FILLED FOR 892 00:43:00,240 --> 00:43:02,760 ALMOST 20 YEARS. 893 00:43:02,760 --> 00:43:13,320 SO WE PRODUCE OR WE HYPOTHESIZED 894 00:43:15,200 --> 00:43:19,320 INITIALLY THAT AP1 CAN 895 00:43:19,320 --> 00:43:20,960 [INDISCERNIBLE], THIS BASED ON 896 00:43:20,960 --> 00:43:25,000 OUR COMPLETION EXPERIMENT SO WE 897 00:43:25,000 --> 00:43:27,160 USE DOUBLE STRAND DNA WITH 898 00:43:27,160 --> 00:43:30,200 SINGLE STRING GAP, SO WE 899 00:43:30,200 --> 00:43:33,800 DESIGNED THIS WITH BIOTIN ON THE 900 00:43:33,800 --> 00:43:36,360 TOP STRAIN, THESE 13 NUCLEAR 901 00:43:36,360 --> 00:43:40,600 TIDE GAPS SO WE ADDED THIS THE 902 00:43:40,600 --> 00:43:42,520 STRUCTURE WE CALL IT SINGLE 903 00:43:42,520 --> 00:43:46,640 STRAND DNA GAP STRUCTURES INTO 904 00:43:46,640 --> 00:43:49,360 HHS, INTERESTINGLY, BOTH THE 13 905 00:43:49,360 --> 00:43:51,720 NUCLEAR SINGLE STRAINED GAP AND 906 00:43:51,720 --> 00:43:54,680 THE OTHER GAP FOR 1 907 00:43:54,680 --> 00:43:57,240 PHOSPHORYLATION, ALSO WE SEE 908 00:43:57,240 --> 00:44:01,600 RECRUITMENT RP COMPLEX, RP 70, 909 00:44:01,600 --> 00:44:03,320 RECRUITMENT FOR THE STRUCTURES, 910 00:44:03,320 --> 00:44:05,680 WE ALSO SEE APE1 RECRUITMENT AS 911 00:44:05,680 --> 00:44:10,240 WELL SO NOT SURPRISINGLY IT WAS 912 00:44:10,240 --> 00:44:11,320 ALL RECOTTERRED, RIGHT. 913 00:44:11,320 --> 00:44:16,520 MORE IMPORTANT WHEN WE REDEPLETE 914 00:44:16,520 --> 00:44:18,960 APE1, AND PHOSPHORYLATION, IT'S 915 00:44:18,960 --> 00:44:23,320 COMPLETELY GONE, AND THAT'S IN 916 00:44:23,320 --> 00:44:27,120 EFFECT RECRUITMENT RPE70 OR NOT 917 00:44:27,120 --> 00:44:29,680 EVEN OLD RPE32, BUT THE 918 00:44:29,680 --> 00:44:30,960 DIFFICULT FIND SUGGEST ATRIP 919 00:44:30,960 --> 00:44:33,120 CANNOT BE RECORDED WITHOUT APE1. 920 00:44:33,120 --> 00:44:35,280 SO THIS IS QUIED INTERESTING 921 00:44:35,280 --> 00:44:36,160 SUGGESTING THAT APE1 COULD BE 922 00:44:36,160 --> 00:44:41,320 THE GUY THAT WE'RE LOOKING FOR 923 00:44:41,320 --> 00:44:43,080 AROUND 20 YEARS. 924 00:44:43,080 --> 00:44:46,680 SO 1 OF THOSE WAS CAN YOU 925 00:44:46,680 --> 00:44:50,560 REVISIT RPA DEPLETION, OKAY, SO 926 00:44:50,560 --> 00:44:52,480 WE DID RPA DEPLETION AND WE 927 00:44:52,480 --> 00:44:56,440 FOUND THAT IT DEPLETED RP A 32, 928 00:44:56,440 --> 00:45:00,040 AND CHECK FOR POSFORALATION BUT 929 00:45:00,040 --> 00:45:01,480 OUR RPA ANTIBODY CUT SOME OF THE 930 00:45:01,480 --> 00:45:06,560 PROTEIN HAD HAS, SO THIS IS COB 931 00:45:06,560 --> 00:45:07,440 CYSTENT WITH PRIOR PUBLICATION 932 00:45:07,440 --> 00:45:08,920 BY KD--SALLY LAWEDDER DALE LAB. 933 00:45:08,920 --> 00:45:13,480 BUT IMPORTANTLY, WITH THAT RPA, 934 00:45:13,480 --> 00:45:17,560 [INDISCERNIBLE] 32, APE 1 935 00:45:17,560 --> 00:45:19,600 BINDING 2 SSDNA GAPS ARE STILL 936 00:45:19,600 --> 00:45:22,320 THERE, AND SOME ARE THE PROTEINS 937 00:45:22,320 --> 00:45:29,240 CAN STILL BE RECRUITED TO THE 938 00:45:29,240 --> 00:45:30,240 SSB [INDISCERNIBLE] STRUCTURES. 939 00:45:30,240 --> 00:45:34,160 SO WE QUANTIFY THE ATRIP ON 940 00:45:34,160 --> 00:45:36,000 Bs, VERSUS WE [INDISCERNIBLE] 941 00:45:36,000 --> 00:45:38,640 ATRIP, WE FOUND YES IT IS 942 00:45:38,640 --> 00:45:41,320 REDUCED BUT VERY MINOR, LIKE 943 00:45:41,320 --> 00:45:41,880 20-30% REDUCTION. 944 00:45:41,880 --> 00:45:47,960 SO THIS COULD BE RPA INDEPENDENT 945 00:45:47,960 --> 00:45:50,120 BUT AE1 DEPENDENT RECRUITMENT OF 946 00:45:50,120 --> 00:45:52,560 OUR ATRIP COMPLEX TO SINGLE 947 00:45:52,560 --> 00:45:54,160 STRAND DNA. 948 00:45:54,160 --> 00:45:57,400 FURTHER MORE, IT'S VERY 949 00:45:57,400 --> 00:46:00,560 INTERESTING WE FIND THAT APE1, 950 00:46:00,560 --> 00:46:06,920 SO THIS IS TAG APE 1, THIS IS 951 00:46:06,920 --> 00:46:08,800 GST, COMBINED TO THE 952 00:46:08,800 --> 00:46:10,080 [INDISCERNIBLE] ATE GUESTS WE 953 00:46:10,080 --> 00:46:12,120 HAVE [INDISCERNIBLE] IN THE 954 00:46:12,120 --> 00:46:14,720 INVITRO SYSTEM, AND THIS APE 955 00:46:14,720 --> 00:46:16,760 INTACT WITH A SINGLE STRAND DNA 956 00:46:16,760 --> 00:46:19,600 IS A LENGTH DEPENDENT, ALSO 957 00:46:19,600 --> 00:46:20,680 CHARACTERIZE THE DOMAIN 958 00:46:20,680 --> 00:46:22,680 REQUIREMENTS WITH THIS, SO IF 959 00:46:22,680 --> 00:46:27,200 YOU COMPLETED THE FIRST 34, THE 960 00:46:27,200 --> 00:46:27,960 MISTRESS [INDISCERNIBLE] ATPE1 961 00:46:27,960 --> 00:46:31,040 YOU LOSE THE BINDING WITH SINGLE 962 00:46:31,040 --> 00:46:31,800 STRAND DNA. 963 00:46:31,800 --> 00:46:33,720 HOWEVER THAT END TERMINAL IS NOT 964 00:46:33,720 --> 00:46:39,920 ENOUGH FOR THIS BINDING. 965 00:46:39,920 --> 00:46:46,120 WE'RE INTERESTED IN APE ALSO 966 00:46:46,120 --> 00:46:47,720 INTACT WITH THE [INDISCERNIBLE] 967 00:46:47,720 --> 00:46:48,800 PROTEIN AND I WANT TO POINT OUT 968 00:46:48,800 --> 00:46:51,920 THAT THE END TERMINAL PROTEIN 969 00:46:51,920 --> 00:46:54,720 DELETION CANNOT INTACT WITH THE 970 00:46:54,720 --> 00:46:56,000 SINGLE STRAND DNA AND 100 IMMUNE 971 00:46:56,000 --> 00:46:58,200 ASSAY DIVISION CANNOT INTACTIVE 972 00:46:58,200 --> 00:46:59,800 WITH THE ATRIP SO WE 973 00:46:59,800 --> 00:47:03,000 CHARACTERIZE THIS 2 TRAG 974 00:47:03,000 --> 00:47:03,320 FRAGMENT APE1. 975 00:47:03,320 --> 00:47:07,320 SO WE WANT TO KNOW WHETHER 976 00:47:07,320 --> 00:47:12,040 APE IN ITSELF CAN RECRUIT TOW 977 00:47:12,040 --> 00:47:13,840 SINGLE STRAIN DNA GAP 978 00:47:13,840 --> 00:47:14,120 STRUCTURES. 979 00:47:14,120 --> 00:47:21,320 YES WHEN WE ADD THE SNA, AND THE 980 00:47:21,320 --> 00:47:25,840 IT CAN BE RECRUITED AND WE 981 00:47:25,840 --> 00:47:27,680 DEPLETED THE FIRST 100 IMMUNO 982 00:47:27,680 --> 00:47:29,600 ASSAYS THAT CANNOT INTACT THE 983 00:47:29,600 --> 00:47:32,800 ATRIP BUT STILL BIND TO SINGLE 984 00:47:32,800 --> 00:47:33,840 STRAND DNA AND CANNOT 985 00:47:33,840 --> 00:47:34,600 [INDISCERNIBLE] ATRIP. 986 00:47:34,600 --> 00:47:36,800 IF YOU COMPLETE THE FORM, CANNOT 987 00:47:36,800 --> 00:47:38,680 INTACT WITH THE SINGLE STRAND 988 00:47:38,680 --> 00:47:40,560 DNA EVEN THOUGH INTACT WITH 989 00:47:40,560 --> 00:47:43,000 [INDISCERNIBLE] ATRIP COOPERATE 990 00:47:43,000 --> 00:47:45,440 BE ABLE TO RECRUIT HIS ATRIP TO 991 00:47:45,440 --> 00:47:47,880 SINGLE STRAND DNA SO THIS IS 992 00:47:47,880 --> 00:47:50,080 STILL THINKING WE EXTRACT, WE 993 00:47:50,080 --> 00:47:52,200 DEPLETE APE1, BUT AT WILD-TYPE 994 00:47:52,200 --> 00:47:54,680 THE MUTANT, THIS LIMITATION 995 00:47:54,680 --> 00:47:58,760 CANNOT RESCUE ATRIP BINDING THIS 996 00:47:58,760 --> 00:47:59,960 MUTATION CANNOTRESS CUE ATRIP 997 00:47:59,960 --> 00:48:00,560 BINDING EITHER. 998 00:48:00,560 --> 00:48:05,760 WE ALSO TESTED THIS IS INDEED 999 00:48:05,760 --> 00:48:08,400 NUCLEUS INDEPENDENT BECAUSE THE 1000 00:48:08,400 --> 00:48:10,880 MUTATION [INDISCERNIBLE] IS INDO 1001 00:48:10,880 --> 00:48:11,720 MUTANT DEFICIENT HAS NOTHING TO 1002 00:48:11,720 --> 00:48:16,400 DID WITH THE RECRUITMENT TO 1003 00:48:16,400 --> 00:48:16,880 SINGLE STRAND DNA. 1004 00:48:16,880 --> 00:48:17,640 MORE INTERESTING AND FURTHER 1005 00:48:17,640 --> 00:48:21,320 MOIRE WE LOOK AT THE APE1 1006 00:48:21,320 --> 00:48:23,400 POTENTIAL INTAKS WITH RPA, WE 1007 00:48:23,400 --> 00:48:26,920 FOUND THE 2 POTENTIAL MOTIF IN 1008 00:48:26,920 --> 00:48:28,400 THE APE1. 1009 00:48:28,400 --> 00:48:34,360 ONE IS THE D69, ANOTHER IS THE 1010 00:48:34,360 --> 00:48:36,120 E48/49 MUTATION CALLED ARM 1011 00:48:36,120 --> 00:48:37,440 BINDING MOTIF 1, ARM BINDING 1012 00:48:37,440 --> 00:48:38,160 MOTIF 2. 1013 00:48:38,160 --> 00:48:40,080 SO A MUTANT EACH OF THEM CAN 1014 00:48:40,080 --> 00:48:45,200 CAUSE SOME SORT OF INTERACTION 1015 00:48:45,200 --> 00:48:46,840 RPEINTERACTION REDUCTION BUT A 1016 00:48:46,840 --> 00:48:49,960 MUTANT WILL DECREASE 1017 00:48:49,960 --> 00:48:50,640 BOTH--INCREASE BOTH 1018 00:48:50,640 --> 00:48:52,160 SIGNIFICANTLY INTACT WITH RPA. 1019 00:48:52,160 --> 00:48:58,040 COME BACK TO THIS RPA MEDIATED 1020 00:48:58,040 --> 00:48:59,920 [INDISCERNIBLE] DOUBLE STRAND 1021 00:48:59,920 --> 00:49:03,040 DNA--YES, IT'S SUFFICIENT TO 1022 00:49:03,040 --> 00:49:04,560 RECRUIT ATRIP IMPORTANT TO 1023 00:49:04,560 --> 00:49:08,760 SINGLE STRAND DNA AND THIS GST 1024 00:49:08,760 --> 00:49:09,960 ALSO CAN DO THE SAME THING BUT 1025 00:49:09,960 --> 00:49:12,000 IF YOU HAVE BOTH, THE 1026 00:49:12,000 --> 00:49:13,560 COMBINATION INDICATE IT'S NOT 1027 00:49:13,560 --> 00:49:16,600 DEPENDENT ON EACH OTHER, IT'S 1028 00:49:16,600 --> 00:49:18,600 NOT SINNER GESTIC SO WE PROPOSE 1029 00:49:18,600 --> 00:49:21,680 THE MODEL IN THE COMTENT OR 1030 00:49:21,680 --> 00:49:23,200 RECONSTITUTION SYSTEM, BOTH RP 1031 00:49:23,200 --> 00:49:28,480 AND ATRIB CAN DO THE RECRUITMENT 1032 00:49:28,480 --> 00:49:32,520 OF ATRIP INDEPENDENTLY, AND 1033 00:49:32,520 --> 00:49:35,520 THERE ARE APUINDEPENDENT IS 1034 00:49:35,520 --> 00:49:35,920 RPEINDEPENDENT AND 1035 00:49:35,920 --> 00:49:36,520 RPEINDEPENDENT. 1036 00:49:36,520 --> 00:49:39,120 SO THAT MEANS ATRIP CAN RECRUIT. 1037 00:49:39,120 --> 00:49:41,480 IT CAN RECRUIT ATRIP TO THE 1038 00:49:41,480 --> 00:49:42,360 SINGLE STRAND DNA. 1039 00:49:42,360 --> 00:49:46,360 HOWEVER, IN THE RP DEPENDENT. 1040 00:49:46,360 --> 00:49:52,840 SOMEHOW AP-1 MAY HAVE TO RELEASE 1041 00:49:52,840 --> 00:49:54,640 INHIBITORY PROTEIN X THAT 1042 00:49:54,640 --> 00:49:56,720 REGULATES RECRUITMENT THROUGH 1043 00:49:56,720 --> 00:49:58,080 THE RPA DEPENDENT MECHANISM. 1044 00:49:58,080 --> 00:50:02,120 SO I WILL GIVE YOU A LITTLE BIT 1045 00:50:02,120 --> 00:50:04,320 OF THE LAST STORY WE RECENTLY 1046 00:50:04,320 --> 00:50:08,960 PUBLISHED ON THE APS FUNCTION IN 1047 00:50:08,960 --> 00:50:10,800 THE [INDISCERNIBLE] AS YOU KNOW 1048 00:50:10,800 --> 00:50:17,240 IN THE NUCLEI, IT'S VERY UNIQUE 1049 00:50:17,240 --> 00:50:20,240 COMPARTMENT WHERE THE RBA IS 1050 00:50:20,240 --> 00:50:23,160 TRANSCRIBED INTO RIBOSOME RNA. 1051 00:50:23,160 --> 00:50:27,040 THIS IS A VERY UNIQUE REGION TO 1052 00:50:27,040 --> 00:50:29,760 FORM [INDISCERNIBLE] THROUGH 1053 00:50:29,760 --> 00:50:31,320 DIFFERENT MULTIPLE 1054 00:50:31,320 --> 00:50:32,480 [INDISCERNIBLE] MECHANISM CALLED 1055 00:50:32,480 --> 00:50:33,680 LIQUID OF LIQUID SEPARATION. 1056 00:50:33,680 --> 00:50:35,680 WE HAVE MARKERS, I WANT TO POINT 1057 00:50:35,680 --> 00:50:39,920 OUT THIS GC COMPONENTS, MPM 1 1058 00:50:39,920 --> 00:50:46,840 INDICATE THE DC COMPONENT OF THE 1059 00:50:46,840 --> 00:50:47,200 NUCLEUS. 1060 00:50:47,200 --> 00:50:49,040 THIS NUCLEI CAN BE STRESSED BY 1061 00:50:49,040 --> 00:50:51,360 DIFFERENT TYPES OF DAMAGE 1062 00:50:51,360 --> 00:50:54,840 INCLUDING DOUBLE STRAND BREAKS, 1063 00:50:54,840 --> 00:50:56,880 INHIBITION, STRESS, OSMATIC 1064 00:50:56,880 --> 00:50:59,880 STRESS, TRIGGER ATN AND ATR DDR 1065 00:50:59,880 --> 00:51:02,040 PATHWAYS AND MORE REEBTLY THE 1066 00:51:02,040 --> 00:51:03,800 APS MEDIATED BY THIS TOPIC P1 1067 00:51:03,800 --> 00:51:09,520 AND OUR STUDY ON APE1 CAN 1068 00:51:09,520 --> 00:51:10,880 ILLUSTRATE THE DIFFERENT 1069 00:51:10,880 --> 00:51:12,760 MECHANISM AND HOW ALL OUR 1070 00:51:12,760 --> 00:51:13,680 PATHWAYS ARE ACTIVATED. 1071 00:51:13,680 --> 00:51:18,520 SO, WHY WE STARTED APE 1 IN APS, 1072 00:51:18,520 --> 00:51:21,560 BECAUSE AP1 HAS THIS MISTRESS 1073 00:51:21,560 --> 00:51:24,320 INTERNAL DOMAIN CALLED THIS IDR 1074 00:51:24,320 --> 00:51:27,080 IN PLACE OF THIS OLDER MOTIF. 1075 00:51:27,080 --> 00:51:30,120 ALSO MEDIATED ARE INTACT BUT 1076 00:51:30,120 --> 00:51:32,160 WHEN WE DID OUR STUDIES IN DEAD 1077 00:51:32,160 --> 00:51:35,280 WHAT WE FIRST TRIED TO SAY 1078 00:51:35,280 --> 00:51:38,120 WHETHER KNOCK DOWN APE1 CAN 1079 00:51:38,120 --> 00:51:42,360 AFFECT H2 TO INDUCE 1080 00:51:42,360 --> 00:51:46,200 PHOSPHORYLATION, THAT'S THE 1081 00:51:46,200 --> 00:51:47,000 CASE. 1082 00:51:47,000 --> 00:51:50,360 WE ALSO USE APETHE BEST 1083 00:51:50,360 --> 00:51:53,240 INHIBITOR MODE CALLED APE 3, 1084 00:51:53,240 --> 00:51:55,320 DOZE DEPENDENT AR3 CAN CAUSE 1085 00:51:55,320 --> 00:51:58,040 REDUCTION OF CHECK 1 1086 00:51:58,040 --> 00:51:58,560 PHOSPHORYLATION, BUT 1087 00:51:58,560 --> 00:52:03,400 [INDISCERNIBLE], THEN MY POST 1088 00:52:03,400 --> 00:52:07,200 DOC DO THE OVEREXPRESSION OF 1089 00:52:07,200 --> 00:52:13,080 YP/AP1 THING AND HE FOUND YFP A 1090 00:52:13,080 --> 00:52:15,720 P1 CAN CAUSE PHOSPHORYLATION 1091 00:52:15,720 --> 00:52:16,400 UPREGULATION BUT SURPRISINGLY HE 1092 00:52:16,400 --> 00:52:21,320 FOUND ALL THE MUTATIONS HE 1093 00:52:21,320 --> 00:52:24,720 TRIED, THE AXO DEFICIENT OR THE 1094 00:52:24,720 --> 00:52:31,440 EDO EXO, OR ALL CAN CAUSE CHECK 1095 00:52:31,440 --> 00:52:33,520 1 PHOSPHORYLATION, AND 1096 00:52:33,520 --> 00:52:33,880 NONCONDITIONS. 1097 00:52:33,880 --> 00:52:37,080 SURPRISE, SO THEN WE TEST 1098 00:52:37,080 --> 00:52:39,400 WHETHER IN THE P A MP1 WHETHER - 1099 00:52:39,400 --> 00:52:42,720 TRACKS WITH THE NUCLEUS, IF WE 1100 00:52:42,720 --> 00:52:44,560 ADDED THIS RECOMBIN ANT PROTEIN, 1101 00:52:44,560 --> 00:52:49,000 WE CAN FORM THIS BI MOLECULE, IF 1102 00:52:49,000 --> 00:52:50,440 WE REMOVE END TERMINAL DOMAIN, 1103 00:52:50,440 --> 00:52:53,160 CANNOT DO IT, WELL, WE ADDED 1104 00:52:53,160 --> 00:52:54,920 THOSE PROTEIN INTO THE SPS 1105 00:52:54,920 --> 00:52:57,840 BUFFER, IT CAN GROW AND FUSE 1106 00:52:57,840 --> 00:53:00,680 INTO A LARGER [INDISCERNIBLE], 1107 00:53:00,680 --> 00:53:02,360 AND THE NUCLEIC EXTRACT WE FOUND 1108 00:53:02,360 --> 00:53:04,280 THIS END TERMINAL DELETION FEELS 1109 00:53:04,280 --> 00:53:05,800 TOAC IV THERAPY POSFORALATION, 1110 00:53:05,800 --> 00:53:09,080 THE PULL DON ASSAY, SHOWS THIS 1111 00:53:09,080 --> 00:53:10,440 END TERMINAL IS VERY IMPORTANT 1112 00:53:10,440 --> 00:53:15,880 FOR INTAKS WITH ATR COMPLEXES 1113 00:53:15,880 --> 00:53:17,040 WITH THE NUCLEIC CONSTRUCTS. 1114 00:53:17,040 --> 00:53:21,080 WE LOOK AT IMAGING OF THE YFAPP, 1115 00:53:21,080 --> 00:53:25,600 WITH THE WILD-TYPE APE1 CAN CO 1116 00:53:25,600 --> 00:53:30,760 LOCALIZE IN THE NUCLEUS AND THE 1117 00:53:30,760 --> 00:53:37,560 BOTH ATR APE1 CO LOCALIZED WITH 1118 00:53:37,560 --> 00:53:38,320 THE YFP 11. 1119 00:53:38,320 --> 00:53:40,960 THOSE IN THE CANCER CELL, BUT WE 1120 00:53:40,960 --> 00:53:43,600 COULDN'T FIND THOSE IN NORMAL 1121 00:53:43,600 --> 00:53:43,920 CELL. 1122 00:53:43,920 --> 00:53:47,360 AS YOU KNOW, THERE ARE 2 1123 00:53:47,360 --> 00:53:49,960 ACTIVATOR TOPICS AND ETA, AND 1 1124 00:53:49,960 --> 00:53:52,240 FOR KINASES, WE FOUND HAVING 1125 00:53:52,240 --> 00:53:59,000 INTERESTING DOMAIN IN APE1 AS IT 1126 00:53:59,000 --> 00:54:01,520 CONSERVE DOMAIN AND THE 1127 00:54:01,520 --> 00:54:02,880 TRANSCRIPT O FAN 9119 1128 00:54:02,880 --> 00:54:08,160 R-MUTATION, NOW ALSO IN THE END 1129 00:54:08,160 --> 00:54:09,560 TERMINAL WHILE I COMPARE THOSE 1130 00:54:09,560 --> 00:54:12,360 RECRUITMENT INTO THE NUCLEUS, WE 1131 00:54:12,360 --> 00:54:14,760 FOUND THIS [INDISCERNIBLE] 7 1132 00:54:14,760 --> 00:54:18,600 MUTATION CANNOT RECRUIT, OR 1133 00:54:18,600 --> 00:54:22,920 CANNOT BE LOCALIZED INTO THE 1134 00:54:22,920 --> 00:54:31,080 NUCLEUS. 1135 00:54:31,080 --> 00:54:32,560 AND THIS [INDISCERNIBLE], THE 7 1136 00:54:32,560 --> 00:54:35,160 R PLURIBUITATION CANNOT TRIGGER 1137 00:54:35,160 --> 00:54:36,320 THE PHOSPHORYLATION, THE TRIP TO 1138 00:54:36,320 --> 00:54:40,160 1 CAN NOW TRIGGER THAT EITHER. 1139 00:54:40,160 --> 00:54:44,160 WE LOOK AT THE BINDING WITH ATR 1140 00:54:44,160 --> 00:54:46,600 AND THE FONT, AND THE R-MUTATION 1141 00:54:46,600 --> 00:54:48,360 VAIM AS WILD-TYPE CAN BIND TO IT 1142 00:54:48,360 --> 00:54:50,800 BUT IT CANNOT FORM THE 1143 00:54:50,800 --> 00:54:54,600 [INDISCERNIBLE] IN THE NUCLEUS 1144 00:54:54,600 --> 00:54:55,040 EXTRACTS. 1145 00:54:55,040 --> 00:54:57,240 MORE IMPORTANT IS HOW WE SEE THE 1146 00:54:57,240 --> 00:54:59,360 KINASES IN THIS, WE USE THE IT 1147 00:54:59,360 --> 00:55:03,960 AS A SUBSTRATE, WE FOUND THIS 1148 00:55:03,960 --> 00:55:11,600 WILD-TYPE APE 1 CHECK 1 BY THIS 1149 00:55:11,600 --> 00:55:13,360 MUTATION KOONLT CHECK 1 BY 1150 00:55:13,360 --> 00:55:17,480 [INDISCERNIBLE] THIS SO INDICATE 1151 00:55:17,480 --> 00:55:22,080 ATR IS VERY ACTIVE IN THE 1152 00:55:22,080 --> 00:55:23,080 KINASES. 1153 00:55:23,080 --> 00:55:25,200 THIS CAN CAUSE THE SYNTHESIS IN 1154 00:55:25,200 --> 00:55:27,560 REDUCTION BUT IT CAN BE REVERSED 1155 00:55:27,560 --> 00:55:28,720 BY ATR INHIBITOR, THE CELL 1156 00:55:28,720 --> 00:55:35,040 ABILITY ALSO CAN BE REDUCED AND 1157 00:55:35,040 --> 00:55:39,080 IT CAN BE REVERSED, THIS IS P53 1158 00:55:39,080 --> 00:55:40,160 POPULATION, SO PUTTING THINGS 1159 00:55:40,160 --> 00:55:43,480 TOGETHER HERE IS THE WORKING 1160 00:55:43,480 --> 00:55:46,760 MODEL THIS IS NOD SURPRISING 1161 00:55:46,760 --> 00:55:48,040 BECAUSE THE APE1 HAS 1162 00:55:48,040 --> 00:55:49,040 [INDISCERNIBLE] ACTIVITY THAT 1163 00:55:49,040 --> 00:55:50,240 ARE IMPORTANT FOR THE HR 1164 00:55:50,240 --> 00:55:57,120 SIGNALING AND THE STRESSFUL 1165 00:55:57,120 --> 00:55:58,640 CONDITIONS SO WE PRESENT OVER 1166 00:55:58,640 --> 00:56:00,800 HERE AND IN CANCER CELLS, 1167 00:56:00,800 --> 00:56:03,280 SOMEHOW THIS CAN FORM THE 1168 00:56:03,280 --> 00:56:04,440 LIQUID, LIQUID SEPARATION, 1169 00:56:04,440 --> 00:56:09,840 THROUGH THEEND TERMINAL, AND THE 1170 00:56:09,840 --> 00:56:13,920 TRANSCRIPT O FAN, AND CAN 1171 00:56:13,920 --> 00:56:16,720 RECRUIT ATR TRIP, AND INTO THE 1172 00:56:16,720 --> 00:56:22,360 NUCLEI, AND ACTIVITY WITH THE 1173 00:56:22,360 --> 00:56:23,120 NUCLEI PATHWAY ACTIVATION EMPLOY 1174 00:56:23,120 --> 00:56:25,480 SO THIS IS A QUICK SUMMARY, THIS 1175 00:56:25,480 --> 00:56:29,000 IS REQUIRES IT TO BE REPAIRED 1176 00:56:29,000 --> 00:56:31,280 FOR SIGNALING, APE1 IS INTACT 1177 00:56:31,280 --> 00:56:35,040 WITH AND RECRUITED TO SINGLE 1178 00:56:35,040 --> 00:56:37,080 STRAND DNA THROUGH 1179 00:56:37,080 --> 00:56:38,080 NON[INDISCERNIBLE] FUNCTION, 1180 00:56:38,080 --> 00:56:43,280 ALSO PRESENT FOR SHOWING IT CAN 1181 00:56:43,280 --> 00:56:44,840 ASSEMBLE BY MITOCHONDRIA, TO 1182 00:56:44,840 --> 00:56:47,840 PROMOTE THE DDR PATHWAY IN 1183 00:56:47,840 --> 00:56:50,240 CANCER BUT NORMAL CELL. 1184 00:56:50,240 --> 00:56:52,280 SO OVERALL APE1 IS ACTIVATOR OF 1185 00:56:52,280 --> 00:56:55,000 THE ATR, DID, DR PATHWAYS. 1186 00:56:55,000 --> 00:56:57,960 SO PUT THEM TOGETHER, BOTH THE 1187 00:56:57,960 --> 00:57:00,600 OLDER AND YOUNGER BROTHER APE1 1188 00:57:00,600 --> 00:57:02,920 AND APE2, IN ADDITION TO THE DNA 1189 00:57:02,920 --> 00:57:06,760 REPAIR FUNCTION, BOTH OF THEM 1190 00:57:06,760 --> 00:57:09,800 ARE ATR, DID, DR IMPORTANT 1191 00:57:09,800 --> 00:57:10,960 CONTROL IN ATR DDR PATHWAY. 1192 00:57:10,960 --> 00:57:15,800 SO YOU CAN AM BACK TO THIS 1193 00:57:15,800 --> 00:57:18,680 SCREEN BY--IN ADDITION TO THE 1194 00:57:18,680 --> 00:57:21,720 BER AND THE MMEJ ACTIVITIES THEY 1195 00:57:21,720 --> 00:57:24,520 FOUND OUT THAT IT HAS SYNTHETIC 1196 00:57:24,520 --> 00:57:26,960 LETHALITY FOR THE BRCA 2 1197 00:57:26,960 --> 00:57:28,960 DEFICIENCIES SO LYLELY ATR, DDR 1198 00:57:28,960 --> 00:57:32,760 DEPOSIT COULD BE 1 OF THE 1199 00:57:32,760 --> 00:57:33,880 MECHANISMS FOR BRCA DEFICIENCY. 1200 00:57:33,880 --> 00:57:39,400 SO LASTLY I WANT TO THANK MY LAB 1201 00:57:39,400 --> 00:57:40,200 MEMBERS AND COLLABORATORS OVER 1202 00:57:40,200 --> 00:57:42,240 THE YEARS AND MY FUNDING AGENCY. 1203 00:57:42,240 --> 00:57:45,360 SO THANK YOU FOR YOUR ATTENTION. 1204 00:57:45,360 --> 00:57:53,720 WE WILL START HERE. 1205 00:57:53,720 --> 00:57:54,720 >> THANK YOU SO MUCH. 1206 00:57:54,720 --> 00:57:56,240 THAT WAS REALLY INTERESTING. 1207 00:57:56,240 --> 00:57:58,280 WE HAVE A NUMBER OF QUESTIONS IN 1208 00:57:58,280 --> 00:58:00,720 THE CHAT, SHOULD I KICK IT OFF, 1209 00:58:00,720 --> 00:58:01,120 KEN? 1210 00:58:01,120 --> 00:58:02,640 >> GO FOR IT. 1211 00:58:02,640 --> 00:58:06,720 >> OKAY, FROM SARAH HINGLE, THIS 1212 00:58:06,720 --> 00:58:09,800 IS IN REFERENCE TO YOUR EARLY 1213 00:58:09,800 --> 00:58:11,200 SLIDES ON APE 2. 1214 00:58:11,200 --> 00:58:13,560 ARE THERE MULTIPLE APE 2S ON 1215 00:58:13,560 --> 00:58:17,480 SINGLE STRANDED DID, NA IN YOUR 1216 00:58:17,480 --> 00:58:19,120 EMPHASIS, THERE ARE MULTIPLE 1217 00:58:19,120 --> 00:58:22,840 BANDS SO I'M CURIOUS WHAT THE 1218 00:58:22,840 --> 00:58:24,000 STOICHIOMETRY IS ON THE 1219 00:58:24,000 --> 00:58:24,760 SUBSTRATE. 1220 00:58:24,760 --> 00:58:27,600 >> SO THE MULTIPLE, CAN REFER 1221 00:58:27,600 --> 00:58:28,240 THE QUESTION AGAIN? 1222 00:58:28,240 --> 00:58:32,480 >> SHE ASKED IF THERE WERE 1223 00:58:32,480 --> 00:58:33,120 MULTIPLE APCEREBELLUMS 2 MEL 1224 00:58:33,120 --> 00:58:35,880 COOLS IN THE STRINGLE STRAND DNA 1225 00:58:35,880 --> 00:58:37,440 EMPHASIS, SINCE THERE WERE 1226 00:58:37,440 --> 00:58:40,200 MULTIPLE BANDS PRODUCED SHE WAS 1227 00:58:40,200 --> 00:58:42,600 WONDERING WHAT THE STOICHIOMETRY 1228 00:58:42,600 --> 00:58:42,760 WAS? 1229 00:58:42,760 --> 00:58:44,200 >> THAT'S A GOOD QUESTION BUT WE 1230 00:58:44,200 --> 00:58:46,040 HAVEN'T FIGURED OUT HOW MANY 1231 00:58:46,040 --> 00:58:47,480 MOLECULES CAN FORM KIND OF A 1 1232 00:58:47,480 --> 00:58:51,040 OR 2 OR MULTIPLE APE TO BINDING 1233 00:58:51,040 --> 00:58:52,480 TO SINGLE STRAND DNA YET. 1234 00:58:52,480 --> 00:58:55,320 IT COULD BE, THAT'S A 1235 00:58:55,320 --> 00:58:57,080 POSSIBILITY BUT WE HAVEN'T 1236 00:58:57,080 --> 00:58:58,600 FIGURED IT OUT YET. 1237 00:58:58,600 --> 00:59:01,720 >> OKAY, SO SARAH ALSO ASKED OUR 1238 00:59:01,720 --> 00:59:03,800 APE2 ASKS IF THEY'RE SENSITIVE 1239 00:59:03,800 --> 00:59:06,800 TO KNOCK OUT INHIBITORS. 1240 00:59:06,800 --> 00:59:07,720 >> COULD GOOD QUESTION, 1 OF 1241 00:59:07,720 --> 00:59:08,440 MANY QUESTIONS. 1242 00:59:08,440 --> 00:59:10,200 WE ARE WORKING ON THAT AND I 1243 00:59:10,200 --> 00:59:15,400 THINK KD--SALLY - 1244 00:59:15,400 --> 00:59:17,160 -[INDISCERNIBLE] IN THE LAB 1245 00:59:17,160 --> 00:59:18,760 IS WORKING ON THAT. 1246 00:59:18,760 --> 00:59:20,360 GOOD QUESTION. 1247 00:59:20,360 --> 00:59:25,160 >> THERE'S A QUESTION FROM 1248 00:59:25,160 --> 00:59:25,640 [INDISCERNIBLE], 1249 00:59:25,640 --> 00:59:26,680 XENOPUSTPRODUCES DOUBLE STRAND 1250 00:59:26,680 --> 00:59:29,080 BREAKS THAT ACTIVATE APM, DO YOU 1251 00:59:29,080 --> 00:59:31,240 THINK WHETHER IT'S ISHT 1252 00:59:31,240 --> 00:59:33,120 INTERACTS WITH APE2 AND STOPS 1253 00:59:33,120 --> 00:59:34,560 STRAND BREAK REPAIR? 1254 00:59:34,560 --> 00:59:41,120 >> YES, GOOD QUESTION AS WELL. 1255 00:59:41,120 --> 00:59:42,360 WE HAVEN'T TEST THIS QUESTION 1256 00:59:42,360 --> 00:59:46,600 DIRECTLY BUT AS YOU KNOW, APE2 1257 00:59:46,600 --> 00:59:52,040 PLACE RULE IN MMEJ PATHWAY, WAIT 1258 00:59:52,040 --> 00:59:54,840 FOR APE TO RECOGNIZE WHERE THE 1259 00:59:54,840 --> 00:59:56,360 DSB ENDS AND PROCESS IT FOR 1260 00:59:56,360 --> 01:00:00,600 REPAIR OR SIGNALING IS UNKNOWN, 1261 01:00:00,600 --> 01:00:02,600 IT'S UNKNOWN. 1262 01:00:02,600 --> 01:00:04,000 HOPEFULLY I CAN TALK TO YOU 1263 01:00:04,000 --> 01:00:08,640 ABOUT IN THIS NEXT COUPLE YEARS 1264 01:00:08,640 --> 01:00:09,680 >> OKAY, DR. SAKAMORO, 1265 01:00:09,680 --> 01:00:11,920 INTERESTING TALK, THANK YOU WHAT 1266 01:00:11,920 --> 01:00:15,360 CANCER ASSOCIATED PROTEIN HELPS 1267 01:00:15,360 --> 01:00:18,320 APE1 RECRUITMENT TO NUCLEI? 1268 01:00:18,320 --> 01:00:20,200 >> THAT'S ANOTHER MILLION 1269 01:00:20,200 --> 01:00:25,000 QUESTION, WE ARE VERY INTERESTED 1270 01:00:25,000 --> 01:00:26,680 IN THIS FINDING. 1271 01:00:26,680 --> 01:00:30,560 WHY IN CANCER CELL WE SAY THIS 1272 01:00:30,560 --> 01:00:32,200 APEOVEREXPRESS CAN BE RECORDED 1273 01:00:32,200 --> 01:00:36,080 TO NUCLEI, BUT NOT NORMAL CELL. 1274 01:00:36,080 --> 01:00:39,120 NOW, WE ARE TESTING APE TO 1275 01:00:39,120 --> 01:00:41,120 RECRUITMENT TO NUCLEI USING 1276 01:00:41,120 --> 01:00:44,200 DIFFERENT CANCER CELL LINES AND 1277 01:00:44,200 --> 01:00:46,040 DIFFERENT NORMAL CELL LINES AND 1278 01:00:46,040 --> 01:00:50,520 TRY TO FIGURE OUT WHAT'S THE 1279 01:00:50,520 --> 01:00:53,080 EXACT UNDERLYING MECHANISM. 1280 01:00:53,080 --> 01:00:55,080 THERE ARE SEVERAL PROPOSED 1281 01:00:55,080 --> 01:00:55,360 NECKANISM. 1282 01:00:55,360 --> 01:00:58,920 FOR EXAMPLE, IS THERE ANY CANCER 1283 01:00:58,920 --> 01:01:01,880 CELL ONCA GENE IN CLUE E I THAT 1284 01:01:01,880 --> 01:01:03,720 CAN RECOGNIZE APE, 1 CAN RECRUIT 1285 01:01:03,720 --> 01:01:06,400 IT OR ANY UNIQUE STRUCTURE IN 1286 01:01:06,400 --> 01:01:12,160 CANCER CELL CAN PROMOTE APE1 TO 1287 01:01:12,160 --> 01:01:12,400 NUCLEI. 1288 01:01:12,400 --> 01:01:13,520 HOPEFULLY NIH WILL GIVE US MORE 1289 01:01:13,520 --> 01:01:17,320 MONEY AND WE CAN FIGURE OUT THE 1290 01:01:17,320 --> 01:01:18,320 DETAILS. 1291 01:01:18,320 --> 01:01:18,560 [LAUGHTER] 1292 01:01:18,560 --> 01:01:18,920 >> ALL RIGHT. 1293 01:01:18,920 --> 01:01:21,680 WE SEE THAT APE 2 BECOMES--THIS 1294 01:01:21,680 --> 01:01:24,560 IS FROM CAROL SHRADER, WE SEE 1295 01:01:24,560 --> 01:01:26,720 THAT APE2 BECOMES 1296 01:01:26,720 --> 01:01:29,960 [INDISCERNIBLE] IN GERMAL LYMPHB 1297 01:01:29,960 --> 01:01:31,240 LYMPHOCYTES WHERE LYMPH1 IS 1298 01:01:31,240 --> 01:01:32,600 GREATLY REDUCED THIS ALTERATION 1299 01:01:32,600 --> 01:01:35,440 OF THE RATIO OF APE1 TO 2 MIGHT 1300 01:01:35,440 --> 01:01:38,840 BE SIMILAR TO YOUR APE2 1301 01:01:38,840 --> 01:01:40,520 TRANSGENIC MOUSE, ALSO APE1 AND 1302 01:01:40,520 --> 01:01:43,400 2 WORK TOGETHER IN B-CELLS 1303 01:01:43,400 --> 01:01:45,280 DURING ANTIBODY'S CLASS SWITCH 1304 01:01:45,280 --> 01:01:46,280 RECOMBINATION, IS ANYTHING KNOWN 1305 01:01:46,280 --> 01:01:49,720 ABOUT THE INTERACTION BETWEEN 1306 01:01:49,720 --> 01:01:50,880 THE 2 BROTHERS? 1307 01:01:50,880 --> 01:01:52,320 >> THAT'S EXCELLENT QUESTION, I 1308 01:01:52,320 --> 01:01:57,000 WOULD SAY, IMMUNE CELLS. 1309 01:01:57,000 --> 01:02:00,440 THERE ARE LOTS OF STUDIES BY 1310 01:02:00,440 --> 01:02:03,640 THEIR GROUP AND I THINK THE 1311 01:02:03,640 --> 01:02:05,320 RATIO--FIRST OF ALL I WANT TO 1312 01:02:05,320 --> 01:02:07,080 SAY THEY MAY DO DIFFERENT. 1313 01:02:07,080 --> 01:02:08,400 THEY HAVE DIFFERENT NUCLEUS 1314 01:02:08,400 --> 01:02:12,520 ACTIVITIES, THE EXPRESSION COULD 1315 01:02:12,520 --> 01:02:15,840 BE DIFFERENT, OUR LAB HAVE SOME 1316 01:02:15,840 --> 01:02:19,480 DATA SHOW ACTUAL INTACT WITH 1317 01:02:19,480 --> 01:02:20,400 EACH OTHER. 1318 01:02:20,400 --> 01:02:21,320 HOW THEY COMPETED WITH EACH 1319 01:02:21,320 --> 01:02:24,640 OTHER, WHO WILL DO MORE, WHO 1320 01:02:24,640 --> 01:02:27,200 WILL DO LESS, DEFINITE DEPENDING 1321 01:02:27,200 --> 01:02:28,920 ON THE CONTACT EITHER IN 1322 01:02:28,920 --> 01:02:30,760 DIFFERENT CELL TYPE, DIFFERENT 1323 01:02:30,760 --> 01:02:33,120 TISSUES AND THERE ARE DIFFERENT 1324 01:02:33,120 --> 01:02:38,120 STRESSFUL CONDITIONS. 1325 01:02:38,120 --> 01:02:40,160 SO MANY POSSIBILITIES. 1326 01:02:40,160 --> 01:02:41,800 THERE'S LESS THAN 100 PAPERS ON 1327 01:02:41,800 --> 01:02:42,160 APE2. 1328 01:02:42,160 --> 01:02:47,200 I HOPE THE NEXT ERA OF APE2 WILL 1329 01:02:47,200 --> 01:02:50,080 BE COME NOTHING THE NEXT 10 1330 01:02:50,080 --> 01:02:50,960 YEARS. 1331 01:02:50,960 --> 01:02:51,720 >> KEN? 1332 01:02:51,720 --> 01:02:52,080 >> THAT'S IT. 1333 01:02:52,080 --> 01:02:53,120 >> OKAY, THERE'S 1 MORE QUESTION 1334 01:02:53,120 --> 01:02:54,720 ACTUALLY THAT GOT ENDED UP BEING 1335 01:02:54,720 --> 01:02:56,440 POSTED TO ME PRIVATELY BUT I 1336 01:02:56,440 --> 01:02:59,040 THINK IT'SA MEANT FOR EVERYBODY, 1337 01:02:59,040 --> 01:03:00,160 IT'S FROM [INDISCERNIBLE] AGAIN 1338 01:03:00,160 --> 01:03:03,320 SAYING GREAT TALKS AND DATA 1339 01:03:03,320 --> 01:03:07,560 THANK, IF APE2 IS UPREGULATED 1340 01:03:07,560 --> 01:03:07,960 AND EXAMINED, WHY? 1341 01:03:07,960 --> 01:03:10,760 DOES IT CORRELATE WITH THE 1342 01:03:10,760 --> 01:03:11,240 DEGREE OF PROLIFERATION? 1343 01:03:11,240 --> 01:03:15,040 AND SECOND QUESTION IS DID THE 1344 01:03:15,040 --> 01:03:16,040 APE2 DRUG REGULATION THE 1345 01:03:16,040 --> 01:03:18,960 SENSITIVE IV THERAPY TUMORS? 1346 01:03:18,960 --> 01:03:20,280 >> WE WOULD LIKE TO BE ABLE TO 1347 01:03:20,280 --> 01:03:21,680 ANSWER THOSE QUESTIONS. 1348 01:03:21,680 --> 01:03:24,480 WE ARE ACTIVELY PURSUE THESE 1349 01:03:24,480 --> 01:03:24,880 QUESTIONS. 1350 01:03:24,880 --> 01:03:30,160 I WOULD SAY VERY LIKELY, BUT I 1351 01:03:30,160 --> 01:03:33,600 HAVEN'T PULL THOSE DATA 1352 01:03:33,600 --> 01:03:33,880 TOGETHER. 1353 01:03:33,880 --> 01:03:36,560 BUT, I WANT TO POINT OUT HERE, 1354 01:03:36,560 --> 01:03:41,640 THE APE2 BIOLOGY AND IN CANCER 1355 01:03:41,640 --> 01:03:42,440 IS VERY COMPLEX. 1356 01:03:42,440 --> 01:03:46,280 OUR RECENT REVIEW ARTICLE IN A. 1357 01:03:46,280 --> 01:03:49,160 E. R. CANCER FOUND THAT APE2 1358 01:03:49,160 --> 01:03:53,320 OVEREXPRESS COULD BE POSITIVE OR 1359 01:03:53,320 --> 01:03:53,920 NETIONATIVELY ASSOCIATED WITH 1360 01:03:53,920 --> 01:03:55,680 SURVIVAL DEPENDING ON THE CANCER 1361 01:03:55,680 --> 01:03:55,880 TYPE. 1362 01:03:55,880 --> 01:04:03,200 SO, IN OTHER WORDS, APECOULD BE 1363 01:04:03,200 --> 01:04:05,400 [INDISCERNIBLE] COULD ALSO BE A 1364 01:04:05,400 --> 01:04:06,880 TUMOR SUPPRESSOR GENE DEPENDING 1365 01:04:06,880 --> 01:04:07,480 ON THE CONTACT. 1366 01:04:07,480 --> 01:04:09,520 SO WE TRY TO FIGURE OUT 1367 01:04:09,520 --> 01:04:11,800 MOLECULAR DETAILS AND REPORT TO 1368 01:04:11,800 --> 01:04:14,040 YOU HOPEFULLY SOON BUT DEPENDING 1369 01:04:14,040 --> 01:04:17,800 ON HOW NIH DECIDE TO FUND US BUT 1370 01:04:17,800 --> 01:04:19,320 I REALLY THANK YOU FOR THE 1371 01:04:19,320 --> 01:04:22,640 CRITICAL QUESTION, FIRST OF ALL 1372 01:04:22,640 --> 01:04:23,360 FOR SURE. 1373 01:04:23,360 --> 01:04:24,240 >> AND THERE'S NO MORE QUESTIONS 1374 01:04:24,240 --> 01:04:27,040 IN THE CHAT BUT THERE IS A 1375 01:04:27,040 --> 01:04:28,760 COMMENT AGAIN FROM CAROL SHRADER 1376 01:04:28,760 --> 01:04:31,280 SAYING I'M GLAD PEOPLE ARE 1377 01:04:31,280 --> 01:04:35,080 FINALLY INTERESTED IN APE2, 1378 01:04:35,080 --> 01:04:37,360 EXCLAMATION, EXCLAMATION, 1379 01:04:37,360 --> 01:04:38,160 EXCLAMATION. 1380 01:04:38,160 --> 01:04:39,640 >> THANKS FOR THE COMMENT AND 1381 01:04:39,640 --> 01:04:40,080 ENCOURAGEMENT. 1382 01:04:40,080 --> 01:04:41,400 >> ALL RIGHT, IF THERE ARE NO 1383 01:04:41,400 --> 01:04:43,160 OTHER QUESTIONS, I'M NOT SEEING 1384 01:04:43,160 --> 01:04:47,800 ANY, IT JUST LEAVES ME TO THANK 1385 01:04:47,800 --> 01:04:49,600 TODAY'S SPEAKER SHAN YAN, THANK 1386 01:04:49,600 --> 01:04:51,400 YOU VERY MUCH FOR A VERY 1387 01:04:51,400 --> 01:04:52,000 INTERESTING TALK. 1388 01:04:52,000 --> 01:04:53,080 OTHER PEOPLE ARE SAYING THANKS 1389 01:04:53,080 --> 01:04:57,640 IN THE CHAT, TOO, SO I ALSO WANT 1390 01:04:57,640 --> 01:04:59,040 TO REMIND EVERYBODY THAT SHAN'S 1391 01:04:59,040 --> 01:05:01,720 TALK WILL SOON BE POSTED ON OUR 1392 01:05:01,720 --> 01:05:04,320 ARCHIVE, I THINK USUALLY WITHIN 1393 01:05:04,320 --> 01:05:08,080 24 HOURS, OUR IT PEOPLE ARE VERY 1394 01:05:08,080 --> 01:05:10,080 GOOD AND ALSO JUST TO WISH YOU 1395 01:05:10,080 --> 01:05:12,120 ALL A HAPPY SUMMER AND SEE YOU 1396 01:05:12,120 --> 01:05:14,080 HERE IN SEPTEMBER FOR JIM 1397 01:05:14,080 --> 01:05:14,320 MORRIS. 1398 01:05:14,320 --> 01:05:16,200 >> THANK YOU ALL FOR YOUR 1399 01:05:16,200 --> 01:05:16,480 ATTENTION. 1400 01:05:16,480 --> 01:05:17,000 >> THANK YOU. 1401 01:05:17,000 --> 00:00:00,000 >> BYE-BYE.