1 00:00:05,120 --> 00:00:06,880 >>WELCOME. 2 00:00:06,880 --> 00:00:09,120 THANK YOU ALL FOR BEING HERE AND 3 00:00:09,120 --> 00:00:15,840 SPECIAL THANKS TO OUR SPEAKER 4 00:00:15,840 --> 00:00:21,080 DR. JUDITH FRYDMAN. 5 00:00:21,080 --> 00:00:24,080 IT IS MY PLEASURE TO WELCOME YOU 6 00:00:24,080 --> 00:00:25,560 AND INTRODUCE THIS WALS LECTURE 7 00:00:25,560 --> 00:00:26,920 TODAY. 8 00:00:26,920 --> 00:00:28,760 NOW SOME INFORMATION ABOUT OUR 9 00:00:28,760 --> 00:00:33,200 SPEAKER. 10 00:00:33,200 --> 00:00:36,280 SHE IS THE DONALD KENNEDY CHAIR 11 00:00:36,280 --> 00:00:38,680 IN THE SCHOOL OF HUMANITIES AND 12 00:00:38,680 --> 00:00:40,760 SCIENCES AND PROFESSOR OF 13 00:00:40,760 --> 00:00:44,080 GENETICS AT STANFORD UNIVERSITY. 14 00:00:44,080 --> 00:00:46,920 AND YOU CAN TELL WHEN HUMANITIES 15 00:00:46,920 --> 00:00:51,520 AND SCIENCES ARE JUXTAPOSED THAT 16 00:00:51,520 --> 00:00:54,400 WE'RE IN FOR A VERY SPECIAL 17 00:00:54,400 --> 00:00:55,040 TREAT TODAY. 18 00:00:55,040 --> 00:00:57,120 THERE IS NO QUESTION. 19 00:00:57,120 --> 00:01:00,480 A LONG-STANDING INTEREST IN 20 00:01:00,480 --> 00:01:02,480 PROTEIN FOLDING AND IN PROTEIN 21 00:01:02,480 --> 00:01:03,680 MISS FOLDING. 22 00:01:03,680 --> 00:01:07,760 HOW IS IT THAT A PROTEIN AS IT 23 00:01:07,760 --> 00:01:11,400 TRAVERSES THROUGH ITS LIFETIME 24 00:01:11,400 --> 00:01:14,040 AS IT GETS PICKED UP AS BEING 25 00:01:14,040 --> 00:01:16,280 READY FOR DISPOSAL HOW IS IT 26 00:01:16,280 --> 00:01:19,520 THAT IT KNOWS HOW TO FOLD? 27 00:01:19,520 --> 00:01:21,880 WHAT HAPPENS WHEN IT MISS FOLDS 28 00:01:21,880 --> 00:01:24,520 BOTH IN THE CELL AT THE 29 00:01:24,520 --> 00:01:26,640 MICROSCOPIC LEVEL AND ALSO TO 30 00:01:26,640 --> 00:01:29,920 INDUCE DISEASE IN ALMOST ANY 31 00:01:29,920 --> 00:01:32,080 ORGAN SYSTEM IN THE BODY. 32 00:01:32,080 --> 00:01:34,720 7 OUR LABORATORY HAS UNCOVERED 33 00:01:34,720 --> 00:01:37,480 BASIC PRINCIPLES OF CHAPERONE 34 00:01:37,480 --> 00:01:40,840 ACTION IN DENOVO PROTEIN FOLDING 35 00:01:40,840 --> 00:01:43,800 AND RECENT WORK FROM HER LAB 36 00:01:43,800 --> 00:01:46,040 SUGGEST THAT PROTEIN MISS 37 00:01:46,040 --> 00:01:50,320 FOLDING PLAYS A ROLE IN NEURO 38 00:01:50,320 --> 00:01:52,640 DEGENERATIVE DISEASES AND IN 39 00:01:52,640 --> 00:01:53,960 AGING THROUGHOUT THE BODY. 40 00:01:53,960 --> 00:01:56,600 THIS REALLY SUGGEST THE 41 00:01:56,600 --> 00:01:58,960 POTENTIAL FOR THERAPEUTIC 42 00:01:58,960 --> 00:02:01,360 STRATEGIES AIMED AT CHAPERONES 43 00:02:01,360 --> 00:02:03,240 AND AT THESE MISS FOLDED 44 00:02:03,240 --> 00:02:07,640 PROTEINS. 45 00:02:07,640 --> 00:02:14,080 DR. FRYDMAN RECEIVED BOTH FROM 46 00:02:14,080 --> 00:02:23,040 THE UNIVERSITY OF BUENOS AIRES. 47 00:02:23,040 --> 00:02:25,480 AND THEN DID A SECOND FELLOWSHIP 48 00:02:25,480 --> 00:02:31,000 AT SHOWN KEPT ERRING INSTITUTE. 49 00:02:31,000 --> 00:02:34,040 SHE MOVED TO STANFORD IN 1996. 50 00:02:34,040 --> 00:02:37,560 AND LITERALLY CLIMBED THE 51 00:02:37,560 --> 00:02:39,680 ACADEMIC LADDER AND BECAME A 52 00:02:39,680 --> 00:02:43,120 FULL PROFESSOR WITH TENURE IN 53 00:02:43,120 --> 00:02:44,560 2010. 54 00:02:44,560 --> 00:02:47,480 SHE HAS EARNED MANY, MANY, MANY 55 00:02:47,480 --> 00:02:53,440 AWARDS AND ACCOLADES INCLUDING 56 00:02:53,440 --> 00:02:55,360 IN 2021 BEING ELECTED TO THE 57 00:02:55,360 --> 00:02:57,080 NATIONAL ACADEMY OF THE 58 00:02:57,080 --> 00:02:57,760 SCIENCES. 59 00:02:57,760 --> 00:03:01,080 SHE IS TRULY PASSIONATE ABOUT 60 00:03:01,080 --> 00:03:01,840 MENTORSHIP. 61 00:03:01,840 --> 00:03:05,280 AND HAS HAD MANY, MANY TRAINEES 62 00:03:05,280 --> 00:03:07,440 IN HER LABORATORY MANY OF WHOM 63 00:03:07,440 --> 00:03:10,160 HAVE BEEN FROM GROUPS THAT ARE 64 00:03:10,160 --> 00:03:13,240 UNDER REPRESENTED IN SCIENCE AND 65 00:03:13,240 --> 00:03:14,840 MEDICINE AND THE PRODUCTIVITY 66 00:03:14,840 --> 00:03:17,480 NOT ONLY OF HER OWN LAB BUT OF 67 00:03:17,480 --> 00:03:20,240 THE LABS THAT HER MENTEES HAVE 68 00:03:20,240 --> 00:03:25,600 ESTABLISHED IS REALLY LEGENDARY. 69 00:03:25,600 --> 00:03:28,360 AND I WOULD LIKE TO GIVE A SHOUT 70 00:03:28,360 --> 00:03:32,920 OUT TO THE COCHAIRS OF THE 71 00:03:32,920 --> 00:03:36,640 PROTECT OHIO STATION US -- GROUP 72 00:03:36,640 --> 00:03:41,120 WHO NOMINATED TODAY'S * SPEAKER 73 00:03:41,120 --> 00:03:42,600 AND ARE RESPONSIBLE FOR HER 74 00:03:42,600 --> 00:03:44,760 COMING TO VISIT US TODAY. 75 00:03:44,760 --> 00:03:51,600 SO WITHOUT FURTHER ADO HER TALK 76 00:03:51,600 --> 00:03:55,560 IS TITLED MOLECULAR ORIGAMI,,THE 77 00:03:55,560 --> 00:03:58,560 DELICATE ART OF PROTEIN FOLDING 78 00:03:58,560 --> 00:04:00,760 AND MISS FOLDING AND ITS 79 00:04:00,760 --> 00:04:03,600 RELEVANCE TO HEALTH AND DISEASE. 80 00:04:03,600 --> 00:04:05,160 WELCOME AND WE'RE LOOKING 81 00:04:05,160 --> 00:04:06,400 FORWARD TO YOUR TALK. 82 00:04:06,400 --> 00:04:11,640 [ APPLAUSE ] 83 00:04:11,640 --> 00:04:13,240 >> THANK YOU SO MUCH FOR THAT 84 00:04:13,240 --> 00:04:16,520 VERY GENEROUS INTRODUCTION. 85 00:04:16,520 --> 00:04:17,440 FREEDMAN. 86 00:04:25,000 --> 00:04:27,760 >>THANK YOU FOR THE INVITATION. 87 00:04:27,760 --> 00:04:29,760 I'M HAVING A WONDERFUL DAY. 88 00:04:29,760 --> 00:04:31,280 REALLY LEARNING A LOT. 89 00:04:31,280 --> 00:04:36,360 IT'S WONDERFUL TO SEE SO MANY 90 00:04:36,360 --> 00:04:38,000 OLD FRIENDS AND COLLEAGUES AND 91 00:04:38,000 --> 00:04:40,960 I'M REALLY HAVING A GREAT TIME. 92 00:04:40,960 --> 00:04:43,440 SO WHAT I HOPE TO DO TODAY IS 93 00:04:43,440 --> 00:04:45,400 GIVE YOU AN OVERVIEW OF THE KIND 94 00:04:45,400 --> 00:04:49,280 OF PROBLEMS AND THE KIND OF WORK 95 00:04:49,280 --> 00:04:51,960 WE DO IN THE LAB TRYING TO 96 00:04:51,960 --> 00:04:54,920 UNDERSTAND HOW PROTEINS FOLD IN 97 00:04:54,920 --> 00:04:57,000 THE CELL AND HOW THE CELL 98 00:04:57,000 --> 00:04:58,440 MANAGES THEIR PROTEIN. 99 00:04:58,440 --> 00:05:01,080 I DON'T KNOW IF YOU SEE MY 100 00:05:01,080 --> 00:05:01,520 CURSOR. 101 00:05:01,520 --> 00:05:10,280 MAYBE I CAN SO WHEN I STARTED TO 102 00:05:10,280 --> 00:05:12,760 BECOME INTERESTED IN HOW 103 00:05:12,760 --> 00:05:14,200 PROTEINS FOLD IN THE CELL OUR 104 00:05:14,200 --> 00:05:15,920 THINKING WAS STRONGLY INFLUENCED 105 00:05:15,920 --> 00:05:19,080 BY THE THINKING OF THE FIELD BY 106 00:05:19,080 --> 00:05:24,000 -- EARLY EXPERIMENTS THAT SHOW 107 00:05:24,000 --> 00:05:26,120 THAT THE INFORMATION FOR THE 108 00:05:26,120 --> 00:05:28,200 UNIQUE THREE DIMENSIONAL 109 00:05:28,200 --> 00:05:31,120 STRUCTURE OF PROTEINS IS SOMEHOW 110 00:05:31,120 --> 00:05:34,400 ENCODED IN IT'S MEAN OH ACID 111 00:05:34,400 --> 00:05:35,720 SEQUENCE. 112 00:05:35,720 --> 00:05:39,840 THE THINK WAS * INPUT WAS NOT 113 00:05:39,840 --> 00:05:41,520 GETTING PROTEINS MADE AND ONCE 114 00:05:41,520 --> 00:05:47,880 THEY WERE MADE IN RIBOSOMES -- 115 00:05:47,880 --> 00:05:51,360 -- I WILL SHOW YOU TODAY 116 00:05:51,360 --> 00:05:53,920 PROBABLY THIS IS PERHAPS THE NOT 117 00:05:53,920 --> 00:05:56,040 TOTALLY GENERALLY APPLICABLE AND 118 00:05:56,040 --> 00:05:57,920 THERE MIGHT BE SOME PROTEINS 119 00:05:57,920 --> 00:06:00,360 THAT ESCAPE THIS GENERALIZATION 120 00:06:00,360 --> 00:06:05,480 BUT EVEN FOR THOSE THAT DO HAVE 121 00:06:05,480 --> 00:06:07,640 -- YOU CAN THINK OF THESE A 122 00:06:07,640 --> 00:06:10,480 LITTLE BIT AS THE JAPANESE ART 123 00:06:10,480 --> 00:06:12,640 OF ORIGAMI WHERE HAVE A SET OF 124 00:06:12,640 --> 00:06:15,080 DETAILED INSTRUCTIONS FOR UNIQUE 125 00:06:15,080 --> 00:06:17,200 THREE DIMENSIONAL SHAPE BUT WHEN 126 00:06:17,200 --> 00:06:20,520 YOU TRY TO DO THEM YOU END UP 127 00:06:20,520 --> 00:06:22,600 WITH A PRODUCT THAT HAS TO BE 128 00:06:22,600 --> 00:06:24,160 SUBJECT TO QUALITY CONTROL AND 129 00:06:24,160 --> 00:06:26,120 WHEN YOU THINK HOW THIS PROCESS 130 00:06:26,120 --> 00:06:27,480 HAPPENS IN THE CELL IT'S NOT 131 00:06:27,480 --> 00:06:30,480 JUST ONE PROTEIN FOLDING BUT 132 00:06:30,480 --> 00:06:33,480 IT'S A HUGE AND VERY DENSE AREA 133 00:06:33,480 --> 00:06:37,600 OF BOTH FOLDED AND FOLDING 134 00:06:37,600 --> 00:06:40,600 PROTEINS SO THE POTENTIAL FOR 135 00:06:40,600 --> 00:06:42,240 PROBLEM MAY BE ACTUALLY VERY 136 00:06:42,240 --> 00:06:43,560 HIGH. 137 00:06:43,560 --> 00:06:47,960 SO KIND OF A WAY TO ENVISION 138 00:06:47,960 --> 00:06:51,160 THIS PROBLEM I OFTEN THINK AND 139 00:06:51,160 --> 00:06:53,040 USE AS AN ILLUSTRATION THIS 140 00:06:53,040 --> 00:06:56,600 STRUCTURE WHICH IS THE STRUCTURE 141 00:06:56,600 --> 00:07:00,520 OF THE COMPLEX SWAN -- 142 00:07:00,520 --> 00:07:02,720 ILLUSTRATES THE COMPLEXITY OF 143 00:07:02,720 --> 00:07:03,160 FOLDING. 144 00:07:03,160 --> 00:07:09,560 THIS IS A NATIVE COMPLEX THAT 145 00:07:09,560 --> 00:07:14,160 HAS TRANSLATED TO MITOCHONDRIAL. 146 00:07:14,160 --> 00:07:16,960 SOME TRANSVERSE MEMBRANES. 147 00:07:16,960 --> 00:07:20,920 OTHERS FOLD INTO THE SOLUBLE 148 00:07:20,920 --> 00:07:21,400 PHASE. 149 00:07:21,400 --> 00:07:25,280 ALL NEED TO MEET AND ASSEMBLE 150 00:07:25,280 --> 00:07:28,720 CORRECTLY AND BEGIN THEIR 151 00:07:28,720 --> 00:07:31,640 CORRECT CO-FACTORS. 152 00:07:31,640 --> 00:07:36,680 WHEN YOU ABOUT ABOUT HOW THESE 153 00:07:36,680 --> 00:07:40,400 ASSEMBLE IT'S MORE DIFFICULT. 154 00:07:40,400 --> 00:07:42,920 BY THE SAME TOKEN YOU CAN ASK 155 00:07:42,920 --> 00:07:47,440 WHAT HAPPENS IF ONE OF THOSE -- 156 00:07:47,440 --> 00:07:52,960 ARE DESTABILIZED. 157 00:07:52,960 --> 00:07:55,200 DOES THIS SUB UNIT GET REMOVED 158 00:07:55,200 --> 00:07:57,680 AND REPLACED WITH A NEW ONE? 159 00:07:57,680 --> 00:08:00,680 DOES THE WHOLE COMPLEX GET 160 00:08:00,680 --> 00:08:01,400 DEGRADED? 161 00:08:01,400 --> 00:08:04,480 HOW ARE THE DECISIONS TO AFFECT 162 00:08:04,480 --> 00:08:06,120 THE QUALITY OF THE PROTEIN 163 00:08:06,120 --> 00:08:06,800 IMPLEMENTED. 164 00:08:06,800 --> 00:08:08,160 THESE ARE THE TYPES OF QUESTIONS 165 00:08:08,160 --> 00:08:10,280 THAT I'M INTERESTED IN MY LAB 166 00:08:10,280 --> 00:08:13,160 AND I THINK THE FIELD IN GENERAL 167 00:08:13,160 --> 00:08:13,920 IS INTERESTED IN. 168 00:08:13,920 --> 00:08:17,440 SO WE WANT TO UNDERSTAND HOW 169 00:08:17,440 --> 00:08:19,200 THEY WORK AND OF COURSE THERE IS 170 00:08:19,200 --> 00:08:20,840 ANOTHER LEVEL OF COMPLEXITY 171 00:08:20,840 --> 00:08:29,040 WHICH IS THAT ALL OF THESE HAS 172 00:08:29,040 --> 00:08:30,720 TO HAPPEN IN THE COMPLEX 173 00:08:30,720 --> 00:08:34,320 ENVIRONMENT OF THE CELL. 174 00:08:34,320 --> 00:08:36,200 SO PERHAPS BECAUSE OF THE 175 00:08:36,200 --> 00:08:37,440 COMPLEXITY OF THE PROBLEM OVER 176 00:08:37,440 --> 00:08:40,880 THE LAST 20 YEARS IT HAS BECOME 177 00:08:40,880 --> 00:08:45,160 APPARENT THAT DEFECTS IN PROTEIN 178 00:08:45,160 --> 00:08:47,160 FOLDING AND QUALITY CONTROL THE 179 00:08:47,160 --> 00:08:52,000 NETWORK OF PROCESSES THAT WE NOW 180 00:08:52,000 --> 00:08:55,120 CALL -- ARE LINKED TO A WIDE 181 00:08:55,120 --> 00:08:57,800 RANGE OF HUMAN DISEASES. 182 00:08:57,800 --> 00:09:00,040 PERHAPS THE MOST FAMOUS ARE THE 183 00:09:00,040 --> 00:09:07,800 AMYLOID DEPOSITS -- CAUSED BY A 184 00:09:07,800 --> 00:09:13,360 CUMULATION OF UNFOLDED SPECIES. 185 00:09:13,360 --> 00:09:14,720 IT'S UNCLEAR ABOUT THE 186 00:09:14,720 --> 00:09:16,360 RELATIONSHIP. 187 00:09:16,360 --> 00:09:21,560 MANY OF THESE DISEASES ARE NEURO 188 00:09:21,560 --> 00:09:24,320 DEGENERATIVE DISEASES BUT THERE 189 00:09:24,320 --> 00:09:33,920 ARE OTHERS THAT ARE LINKED SUCH. 190 00:09:33,920 --> 00:09:35,400 -- LINKED. 191 00:09:35,400 --> 00:09:37,880 8 THERE ARE ALSO PROTEIN FOLDING 192 00:09:37,880 --> 00:09:40,080 DEFECTS THAT ARISE FROM 193 00:09:40,080 --> 00:09:41,920 MUTATIONS AND GENERALLY THESE 194 00:09:41,920 --> 00:09:44,480 LEAD NOT TO ACCUMULATION OF MISS 195 00:09:44,480 --> 00:09:48,000 FOLDED PROTEIN BUT TO A LOSS OF 196 00:09:48,000 --> 00:09:49,280 FUNCTION. 197 00:09:49,280 --> 00:09:51,760 PERHAPS THE BEST STUDY IS THE 198 00:09:51,760 --> 00:09:55,640 MUTATION THAT LEADS TO CYSTIC 199 00:09:55,640 --> 00:09:57,440 FIBROSIS BUT THERE ARE MANY 200 00:09:57,440 --> 00:10:01,040 OTHER MUTATIONS LINKED TO CANCER 201 00:10:01,040 --> 00:10:02,000 ETC. 202 00:10:02,000 --> 00:10:05,200 AND OF COURSE BECAUSE THE NATURE 203 00:10:05,200 --> 00:10:08,280 OF PROTEINS'S ABILITIES SO 204 00:10:08,280 --> 00:10:10,760 DELICATE AND AS THEY SAY 205 00:10:10,760 --> 00:10:14,080 MARGINALLY STABLE A NUMBER OF 206 00:10:14,080 --> 00:10:18,560 STRESSES -- CAUSE THIS HAVE BEEN 207 00:10:18,560 --> 00:10:22,120 LINKED TO A NUMBER OF 208 00:10:22,120 --> 00:10:23,040 PATHOLOGICAL STATES INCLUDING 209 00:10:23,040 --> 00:10:24,080 STROKES. 210 00:10:24,080 --> 00:10:27,280 7 AND I INCLUDE HERE -- THIS IS 211 00:10:27,280 --> 00:10:29,760 NOT A DISEASE BUT PERHAPS A 212 00:10:29,760 --> 00:10:31,640 DISEASE WE ALL WANT TO HAVE BUT 213 00:10:31,640 --> 00:10:34,040 IT IS LINKED TO A LARGE NUMBER 214 00:10:34,040 --> 00:10:36,400 OF HUMAN PATHOLOGIES AND THERE 215 00:10:36,400 --> 00:10:38,600 IS INCREASING EVIDENCE THAT THE 216 00:10:38,600 --> 00:10:41,240 CAPACITY OF CELLS THAT MAINTAIN 217 00:10:41,240 --> 00:10:44,480 PROTECT OHIO STATION US * AND 218 00:10:44,480 --> 00:10:47,000 THIS IS ONE OF THE REASONS OR 219 00:10:47,000 --> 00:10:52,200 THE FACTORS THAT ARE LINKED TO 220 00:10:52,200 --> 00:10:54,360 GREAT INCIDENCES OF DISEASES 221 00:10:54,360 --> 00:10:55,920 THAT OCCUR WITH AGING. 222 00:10:55,920 --> 00:10:58,960 IN A VERY, VERY SIMPLISTIC 223 00:10:58,960 --> 00:11:02,400 MANNER WE CAN THINK OF PRO OHIO 224 00:11:02,400 --> 00:11:04,480 STATION US AS THE MULTIPLE FATES 225 00:11:04,480 --> 00:11:06,240 THAT HAPPEN TO A PROTEIN IN THE 226 00:11:06,240 --> 00:11:16,360 CELL. 227 00:11:20,120 --> 00:11:20,560 *. 228 00:11:20,560 --> 00:11:24,320 WE KNOW THAT MANY CHAP REASONS 229 00:11:24,320 --> 00:11:25,720 FACILITATE THIS PROCESS. 230 00:11:25,720 --> 00:11:27,720 IT HAS TO BE MONITORED FOR 231 00:11:27,720 --> 00:11:29,360 INTAKE TREE AND ALSO INVOLVED IN 232 00:11:29,360 --> 00:11:31,000 THE PROCESS. 233 00:11:31,000 --> 00:11:35,240 ALSO INVOLVED IN RESCUING OR 234 00:11:35,240 --> 00:11:37,040 STABILIZING AND ONCE A PROTEIN 235 00:11:37,040 --> 00:11:39,840 IS IN THE CELL BUT IS NON-NATIVE 236 00:11:39,840 --> 00:11:41,840 A DECISION HAS TO BE TAKEN 237 00:11:41,840 --> 00:11:44,040 WHETHER TO REFOLD IT OR SEND IT 238 00:11:44,040 --> 00:11:47,560 FOR CLEARANCE THROUGH A NUMBER 239 00:11:47,560 --> 00:11:50,400 OF DEGRADATION PATHWAYS. 240 00:11:50,400 --> 00:11:55,040 SO THE PROTEIN IS STARTED AND 241 00:11:55,040 --> 00:11:57,600 DEGRADED AND IF IT FAILS YOU 242 00:11:57,600 --> 00:12:00,920 START TO ACCUMULATE MISS FOLDED 243 00:12:00,920 --> 00:12:02,280 SPECIES INCLUDING AMYLOID 244 00:12:02,280 --> 00:12:04,200 AGGREGATES AND WE DON'T 245 00:12:04,200 --> 00:12:07,040 UNDERSTAND HOW THIS DIFFERENT 246 00:12:07,040 --> 00:12:09,280 PHASE OF THE PROTEINS ARE 247 00:12:09,280 --> 00:12:10,200 COORDINATED. 248 00:12:10,200 --> 00:12:12,760 BECAUSE PROTEINS ARE COMPLEX AND 249 00:12:12,760 --> 00:12:15,200 THE CELLULAR FATES ARE VERY 250 00:12:15,200 --> 00:12:17,640 COMPLEX THE NETWORK THAT DEALS 251 00:12:17,640 --> 00:12:20,160 WITH THESE PROBLEMS IS ALSO VERY 252 00:12:20,160 --> 00:12:20,960 COMPLEX. 253 00:12:20,960 --> 00:12:22,760 I TOOK THIS PICTURE OUT OF A 254 00:12:22,760 --> 00:12:24,720 CATALOG AND WHAT I WANT TO BRING 255 00:12:24,720 --> 00:12:27,880 HOME WITH THIS PICTURE IS THE 256 00:12:27,880 --> 00:12:30,120 MULTIPLICITY OF DIFFERENT 257 00:12:30,120 --> 00:12:32,240 STRUCTURAL AND MECHANISTIC 258 00:12:32,240 --> 00:12:35,400 DIVERSITY THERE IS IN THIS 259 00:12:35,400 --> 00:12:37,960 MACHINERY. 260 00:12:37,960 --> 00:12:39,680 THE MACHINERY THERE ARE MANY 261 00:12:39,680 --> 00:12:45,760 PEOPLE THAT WORK ON THESE -- 262 00:12:45,760 --> 00:12:47,200 THAT FUNCTION WITH THE UNIVERSE 263 00:12:47,200 --> 00:12:49,520 OF DIFFERENT CO-FACTORS THAT 264 00:12:49,520 --> 00:12:52,040 REGULATE SUB STRAIGHT BINDING 265 00:12:52,040 --> 00:12:52,640 AND ACTIVITY. 266 00:12:52,640 --> 00:12:54,640 THERE ARE OTHER TYPES OF VERY 267 00:12:54,640 --> 00:12:57,680 LARGE MACHINES SUCH AS THE 268 00:12:57,680 --> 00:13:01,720 CHAPERONES THAT ARE ALSO 269 00:13:01,720 --> 00:13:08,600 DEPENDENT. 270 00:13:08,600 --> 00:13:10,560 -- AND OTHER CO-FACTORS. 271 00:13:10,560 --> 00:13:12,880 7 WE DON'T UNDERSTAND HOW THIS 272 00:13:12,880 --> 00:13:14,320 MINNESOTA MACHINERY WORKS 273 00:13:14,320 --> 00:13:14,960 TOGETHER. 274 00:13:14,960 --> 00:13:18,080 ONE THING ALL CHAPERONES HAVE IN 275 00:13:18,080 --> 00:13:20,200 COMMON IS THEY RECOGNIZE NO 276 00:13:20,200 --> 00:13:21,840 NATIVE PROTEINS BUT BECAUSE OF 277 00:13:21,840 --> 00:13:24,720 THEIR DIFFERENT STRUCTURE AND 278 00:13:24,720 --> 00:13:26,280 MECHANISM EACH TYPE WILL DO 279 00:13:26,280 --> 00:13:28,000 SOMETHING DIFFERENT TO THE 280 00:13:28,000 --> 00:13:30,080 NATIVE PROTEIN. 281 00:13:30,080 --> 00:13:32,280 SO A LITTLE TAKE HOME MESSAGE OF 282 00:13:32,280 --> 00:13:34,840 WHAT I'VE TOLD YOU SO FAR IS THE 283 00:13:34,840 --> 00:13:39,560 NETWORK IS VERY COMPLEX AND THIS 284 00:13:39,560 --> 00:13:41,360 FUNCTION IS LINKED TO A WIDE 285 00:13:41,360 --> 00:13:42,360 RANGE OF DISEASES. 286 00:13:42,360 --> 00:13:44,360 I'M GOING TO GIVE YOU AN 287 00:13:44,360 --> 00:13:46,480 OVERVIEW OF WORK IN MY LAB 288 00:13:46,480 --> 00:13:49,120 TRYING TO UNDERSTAND DIFFERENT 289 00:13:49,120 --> 00:13:50,600 ASPECTS OF THE MACHINERY. 290 00:13:50,600 --> 00:13:51,880 MY FIRST INTEREST AND WHEN I 291 00:13:51,880 --> 00:13:54,800 STARTED TO WORK ON THIS PROBLEM 292 00:13:54,800 --> 00:13:56,480 IS WHAT HAPPENS WHEN A PROTEIN 293 00:13:56,480 --> 00:13:58,040 IS TRANSLATED? 294 00:13:58,040 --> 00:14:00,720 SO AT THAT TIME MOST OF THE 295 00:14:00,720 --> 00:14:03,560 PARADIGMS OF FOLDING WERE KIND 296 00:14:03,560 --> 00:14:06,080 OF A NUANCE IN STYLE OF 297 00:14:06,080 --> 00:14:08,640 EXPERIMENT MEANING A PROTEIN WAS 298 00:14:08,640 --> 00:14:10,880 FULLY MADE PROTEIN WAS DENATURED 299 00:14:10,880 --> 00:14:14,120 AND THEN REFOLDED IN THE 300 00:14:14,120 --> 00:14:16,120 PRESENCE OF CHAP REASON. 301 00:14:16,120 --> 00:14:20,120 BUT I WAS INTERESTED IN HOW THE 302 00:14:20,120 --> 00:14:22,680 CONSTRAINTS OF TRANSLATION -- 303 00:14:22,680 --> 00:14:25,160 PRESENTATION OF THE PEPTIDE TO 304 00:14:25,160 --> 00:14:29,480 THE CYTOPLASM AFFECTED FOLDING. 305 00:14:29,480 --> 00:14:30,840 THIS ESTABLISHED TWO PRINCIPLES 306 00:14:30,840 --> 00:14:32,240 THAT WE'RE STILL TRYING TO 307 00:14:32,240 --> 00:14:32,880 UNDERSTAND. 308 00:14:32,880 --> 00:14:37,240 THE FIRST ONE IS THAT PROTEINS 309 00:14:37,240 --> 00:14:40,680 START TO FOLD AND EVOLVE TO FOLD 310 00:14:40,680 --> 00:14:43,080 IN A DOMAIN WEISMANER AND 311 00:14:43,080 --> 00:14:47,280 SECONDLY CHAPERONES WERE 312 00:14:47,280 --> 00:14:49,880 RECRUITED TO THE NASAN. 313 00:14:49,880 --> 00:14:53,200 AND WHAT WE FOUND IS THAT 314 00:14:53,200 --> 00:14:54,680 CHAPERONES SEEM TO BE RECRUITED 315 00:14:54,680 --> 00:14:57,000 IN A SPECIFIC MANNER SUGGESTING 316 00:14:57,000 --> 00:15:00,560 THERE MUST HAVE BEEN -- CROSS 317 00:15:00,560 --> 00:15:05,440 TALK. 318 00:15:05,440 --> 00:15:07,120 OVER THE YEARS WE'VE LEARNED A 319 00:15:07,120 --> 00:15:08,680 LOT ABOUT THIS MACHINERY. 320 00:15:08,680 --> 00:15:13,800 WE'VE DONE A LOT OF WORK. 321 00:15:13,800 --> 00:15:15,520 TO UNDERSTAND THE PROCESSES. 322 00:15:15,520 --> 00:15:21,800 ONE OF THE FIRST THINGS WAS WORK 323 00:15:21,800 --> 00:15:26,400 WHERE IT WAS LOOKED AT THE 324 00:15:26,400 --> 00:15:28,640 RELATION -- IN RESPONSE TO 325 00:15:28,640 --> 00:15:30,640 STRESS AND THIS WORK LED HER TO 326 00:15:30,640 --> 00:15:33,840 FIND THAT THERE WERE TWO 327 00:15:33,840 --> 00:15:35,280 DISTINCT NETWORKS. 328 00:15:35,280 --> 00:15:42,520 ONE THAT WAS TRANSCRIPTIONALLY 329 00:15:42,520 --> 00:15:49,080 CO-REGULATED AND ARE INVOLVED IN 330 00:15:49,080 --> 00:15:51,640 NEWLY MADE PROTEINS. 331 00:15:51,640 --> 00:15:54,320 AND THE OTHER ONE WAS A NETWORK 332 00:15:54,320 --> 00:15:56,960 OF CHAPERONES THAT WAS INDUCED 333 00:15:56,960 --> 00:15:59,160 BY STRESS AND MANY OF THEM ARE 334 00:15:59,160 --> 00:16:00,400 WELL-KNOWN PROTEINS THAT ARE 335 00:16:00,400 --> 00:16:04,600 INVOLVED IN RESCUING THE 336 00:16:04,600 --> 00:16:05,560 PROTEINS DURING STRESS. 337 00:16:05,560 --> 00:16:08,080 OVER THE YEARS MY LAB AND MANY 338 00:16:08,080 --> 00:16:10,000 OTHER LABS HAVE DEFINED A NUMBER 339 00:16:10,000 --> 00:16:14,840 OF CHAP REASONS THAT INTERACT 340 00:16:14,840 --> 00:16:19,320 PHYSICALLY WITH THE RIBOSOME 341 00:16:19,320 --> 00:16:25,240 FACTOR. 342 00:16:25,240 --> 00:16:28,600 AS WELL AS OTHER CO-FACTORS. 343 00:16:28,600 --> 00:16:30,040 WHEN ONE OF THE FIRST THINGS 344 00:16:30,040 --> 00:16:33,680 THAT KIND OF LIKE -- WAS 345 00:16:33,680 --> 00:16:34,720 PUZZLING AT THE TIME AND YOU 346 00:16:34,720 --> 00:16:36,600 KNOW WE'RE STILL TRYING TO 347 00:16:36,600 --> 00:16:42,880 UNDERSTAND IS HOW DOES THE NASAN 348 00:16:42,880 --> 00:16:43,920 SELECT? 349 00:16:43,920 --> 00:16:45,840 SO ONE POSSIBILITIES THAT THEY 350 00:16:45,840 --> 00:16:50,240 CAN ALL INTERACT AND COMPETE FOR 351 00:16:50,240 --> 00:16:51,680 BINDING TO THE CHAIN BUT OVER 352 00:16:51,680 --> 00:16:53,240 THE YEARS WORK FROM MY LAB 353 00:16:53,240 --> 00:17:02,800 SHOWED THERE IS DISTINCT SPEAKS 354 00:17:02,800 --> 00:17:09,120 I SEE -- * A SPECIFIC SUBSTANCE 355 00:17:09,120 --> 00:17:10,160 OF PEPTIDES WITH SPECIFIC 356 00:17:10,160 --> 00:17:19,960 DOMAINS. 357 00:17:19,960 --> 00:17:22,520 THIS LEADS TO ANOTHER QUESTION. 358 00:17:22,520 --> 00:17:25,960 HOW DO YOU MAKE THE RIGHT 359 00:17:25,960 --> 00:17:29,360 DECISION AND WHAT DETERMINES THE 360 00:17:29,360 --> 00:17:32,320 SPECIFICITY. 361 00:17:32,320 --> 00:17:34,920 YOU HAVE A CHAIN EMERGING FROM 362 00:17:34,920 --> 00:17:36,920 THE SITE OH PLASM. 363 00:17:36,920 --> 00:17:42,520 THERE ARE * -- AND OTHER 364 00:17:42,520 --> 00:17:44,600 FACTORS. 365 00:17:44,600 --> 00:17:48,760 AND THEN YOU'VE MERGING FROM THE 366 00:17:48,760 --> 00:17:52,480 RIBOSOME AN INCOMPLETE PROTEIN. 367 00:17:52,480 --> 00:17:54,840 AND THIS HAS TO BE INTERPRETED 368 00:17:54,840 --> 00:17:58,680 BY THIS SYSTEM OF EITHER 369 00:17:58,680 --> 00:18:00,720 MODIFYING ENZYMES. 370 00:18:00,720 --> 00:18:05,160 SO IF YOU THINK ABOUT TARGETING 371 00:18:05,160 --> 00:18:08,800 AND YOU CAN SEE THIS INCOMPLETE 372 00:18:08,800 --> 00:18:11,560 MOTIF -- IF YOU JUST WAIT A 373 00:18:11,560 --> 00:18:13,920 COUPLE OF SECONDS THESE ACTUALLY 374 00:18:13,920 --> 00:18:19,800 IS AN SRP BINDING MOTIF. 375 00:18:19,800 --> 00:18:22,840 SO * THERE MIGHT BE SOME 376 00:18:22,840 --> 00:18:28,680 MISTAKES MADE OR AN EQUILIBRIUM 377 00:18:28,680 --> 00:18:32,200 -- HOWEVER WORK FROM PHYLLIS IN 378 00:18:32,200 --> 00:18:38,720 MY LAB SHOWS THAT THERE IS 379 00:18:38,720 --> 00:18:39,800 SPECIFICITY. 380 00:18:39,800 --> 00:18:41,800 SO ONE OF THE THINGS THAT WE'RE 381 00:18:41,800 --> 00:18:46,360 TRYING TO UNDERSTAND IS HOW DO 382 00:18:46,360 --> 00:18:54,640 THESE IN -- ENSURE. 383 00:18:54,640 --> 00:18:57,240 I'M GOING TO GIVE YOU A SUMMARY 384 00:18:57,240 --> 00:18:59,760 AND THEN I'M HAPPY TO TELL YOU 385 00:18:59,760 --> 00:19:01,560 MORE ABOUT THIS IF ANYONE IS 386 00:19:01,560 --> 00:19:01,960 INTERESTED. 387 00:19:01,960 --> 00:19:04,800 HOW WE'RE TRYING TO STUDY THE 388 00:19:04,800 --> 00:19:05,040 PROCESS. 389 00:19:05,040 --> 00:19:09,120 WE HAVE FOUND THAT BOTH HBS70 390 00:19:09,120 --> 00:19:13,200 AND THE CHAPERONE -- INVITRO 391 00:19:13,200 --> 00:19:15,240 THEY CAN COMPETE FOR SUB STRAITS 392 00:19:15,240 --> 00:19:18,160 BUT WE KNOW THAT INVITRO THEY 393 00:19:18,160 --> 00:19:23,200 INTERACT WITH DIFFERENT REGIONS 394 00:19:23,200 --> 00:19:28,440 OF THE POLLY PEPTIDE. 395 00:19:28,440 --> 00:19:32,040 * AND WE USE SELECTIVE RIVALS TO 396 00:19:32,040 --> 00:19:34,240 IDENTIFY WHEN THIS DIFFERENT 397 00:19:34,240 --> 00:19:36,560 CHAPERONS ARE RECRUITED TO THE 398 00:19:36,560 --> 00:19:39,000 CHAIN AND WHEN THEY COME AFTER 399 00:19:39,000 --> 00:19:39,880 THE CHAIN. 400 00:19:39,880 --> 00:19:41,720 ONE EXAMPLE FROM A PAPER 401 00:19:41,720 --> 00:19:43,880 PUBLISHED A COUPLE OF YEARS AGO 402 00:19:43,880 --> 00:19:46,800 BY KEVIN STEIN SHOWING WHERE -- 403 00:19:46,800 --> 00:19:48,760 BINDS IN RELATIONSHIP TO THE 404 00:19:48,760 --> 00:19:51,600 DOMAINS EMERGING FROM THE 405 00:19:51,600 --> 00:19:54,240 RIBOSOME AND YOU CAN SEE THEY 406 00:19:54,240 --> 00:19:55,960 HAVE DIFFERENT BINDING SITES. 407 00:19:55,960 --> 00:19:57,480 THEY RECOGNIZE THAT THE DOMAINS 408 00:19:57,480 --> 00:19:59,360 HAVE DIFFERENT POINTS WHEN THEY 409 00:19:59,360 --> 00:20:03,560 COME OUT OF THE RIBOSOME AND 410 00:20:03,560 --> 00:20:05,360 WHEN WE MAP SPECIFIC STRUCTURES 411 00:20:05,360 --> 00:20:09,080 OF PROTEINS AND PROTEIN DOMAINS 412 00:20:09,080 --> 00:20:12,960 WE SEE THAT IT BINDS IN SPECIFIC 413 00:20:12,960 --> 00:20:16,880 REGIONS OF THE SUB DOMAINS OF 414 00:20:16,880 --> 00:20:19,920 THIS PARTICULAR -- BLADE. 415 00:20:19,920 --> 00:20:21,920 AND WE'RE TRYING TO UNDERSTAND 416 00:20:21,920 --> 00:20:26,400 HOW THIS IS REGULATED AND HOW 417 00:20:26,400 --> 00:20:29,560 THIS INFLUENCES FOLDING. 418 00:20:29,560 --> 00:20:32,800 THE EXPERIMENTS THAT WE TRY TO 419 00:20:32,800 --> 00:20:35,560 UNDERSTAND -- HOW IT HAPPENS 420 00:20:35,560 --> 00:20:40,480 DURING TRANSLATION AND AT THE 421 00:20:40,480 --> 00:20:40,760 RIBOSOME. 422 00:20:40,760 --> 00:20:46,000 THERE ARE SOME PROTEINS TARGETED 423 00:20:46,000 --> 00:20:50,320 BY SRP. 424 00:20:50,320 --> 00:20:52,720 IT RECOGNIZES IT EARLY ON WHILE 425 00:20:52,720 --> 00:20:56,520 STILL IN THE TUNNEL. 426 00:20:56,520 --> 00:20:59,880 THEY BIND TO DISTINCT REGIONS 427 00:20:59,880 --> 00:21:03,440 AND HAVE DISTINCT SPECIFICITY 428 00:21:03,440 --> 00:21:07,960 BUT SOME ROW TEENS NEED TO HAVE 429 00:21:07,960 --> 00:21:12,000 HIERARCHICAL INTERACTION. 430 00:21:12,000 --> 00:21:15,280 OBVIOUSLY THE SPECIFICITY OF 431 00:21:15,280 --> 00:21:18,800 CHAPERONES IS REGULATED ALSO BY 432 00:21:18,800 --> 00:21:25,960 FOLDING AND WE' WE ARE INTERESTN 433 00:21:25,960 --> 00:21:28,960 UNDERSTANDING. 434 00:21:28,960 --> 00:21:31,760 NOW ALL OF THESE STUDIES ALSO 435 00:21:31,760 --> 00:21:33,880 PROVIDED ONE SURPRISE THAT I'M 436 00:21:33,880 --> 00:21:35,640 GOING TO COME BACK LATER IN THE 437 00:21:35,640 --> 00:21:36,120 TALK. 438 00:21:36,120 --> 00:21:40,480 WHICH IS THAT WE FOUND THAT 439 00:21:40,480 --> 00:21:44,040 ELONGATION RATE. 440 00:21:44,040 --> 00:21:48,320 THE RHYTHM -- IS ACTUALLY PRO 441 00:21:48,320 --> 00:21:49,800 FUNDLY INFLUENTIAL TO THE FATE 442 00:21:49,800 --> 00:21:51,840 OF THE POLLY PEPTIDE. 443 00:21:51,840 --> 00:21:54,880 SO THE FIRST ONE TO NOTICE THIS 444 00:21:54,880 --> 00:21:58,000 WAS SEBASTIAN -- THAT FOUND THAT 445 00:21:58,000 --> 00:22:00,600 THERE WERE PATTERNS OF OPTIMAL 446 00:22:00,600 --> 00:22:09,200 AND NONOPTIMAL -- ---AND TURNS 447 00:22:09,200 --> 00:22:15,480 AND LOOPS CONNECTING DOMAINS AND 448 00:22:15,480 --> 00:22:17,800 THEN THEY SHOWED THAT THE EARLY 449 00:22:17,800 --> 00:22:23,160 RECOGNITION OF SRP WAS ALSO 450 00:22:23,160 --> 00:22:25,280 PROMOTED BY LOCAL SLOW DOWN 451 00:22:25,280 --> 00:22:27,480 REGIONS THAT FACILITATED 452 00:22:27,480 --> 00:22:29,280 INTERACTION OF SRP. 453 00:22:29,280 --> 00:22:33,960 8 AND FINALLY KEVIN STEIN FOUND 454 00:22:33,960 --> 00:22:38,640 THAT BOTH HPS70 -- BOUND AT 455 00:22:38,640 --> 00:22:48,320 REGIONS THAT ARE DETERMINED. 456 00:22:48,320 --> 00:22:52,280 -- THIS TAKES THE PARADIGM 457 00:22:52,280 --> 00:22:57,360 FURTHER AND THE MESSAGE AND THE 458 00:22:57,360 --> 00:22:59,840 SEQUENCE ALSO CO-EVOLVED TO 459 00:22:59,840 --> 00:23:02,800 ACCOMMODATE OR TO ATTUNED THE 460 00:23:02,800 --> 00:23:07,760 LONG GOINGS * RATE TO PROMOTE 461 00:23:07,760 --> 00:23:10,520 OPTIMAL FOLDING OR ULTIMATE FATE 462 00:23:10,520 --> 00:23:12,240 OF THAT CHAIN. 463 00:23:12,240 --> 00:23:18,520 THIS SUGGEST THATY LONG GOINGS 464 00:23:18,520 --> 00:23:29,120 -- * I'LL COME BACK TO THIS AT E 465 00:23:35,720 --> 00:23:37,120 END OF MY TALK 466 00:23:37,120 --> 00:23:38,800 THE OTHER QUESTION THAT WE WANT 467 00:23:38,800 --> 00:23:41,920 TO UNDERSTAND IS HOW CHAPERONES 468 00:23:41,920 --> 00:23:43,760 WORK. 469 00:23:43,760 --> 00:23:49,360 WHAT DO 10% OF THE PROTEINS 470 00:23:49,360 --> 00:23:52,560 REQUIRE HSP70 TO FOLD? 471 00:23:52,560 --> 00:23:56,200 AND ONE OF THE MAIN FOCUSES IS 472 00:23:56,200 --> 00:23:58,840 TO UNDERSTAND THE MECHANISM OF 473 00:23:58,840 --> 00:24:00,680 THIS RING SHAPE CHAPERONE. 474 00:24:00,680 --> 00:24:04,080 AVENUE YOU CAN SEE A SIDE VIEW 475 00:24:04,080 --> 00:24:05,880 OF THE STRUCTURE. 476 00:24:05,880 --> 00:24:16,320 AND THIS IS THE TOP VIEW. 477 00:24:17,560 --> 00:24:21,680 -- AND IF YOU TURN IT 90 DEGREES 478 00:24:21,680 --> 00:24:24,400 YOU CAN SEE THAT EACH RING HAS A 479 00:24:24,400 --> 00:24:28,440 CENTRAL CHAMBER THAT HAS A LEGAL 480 00:24:28,440 --> 00:24:28,640 GATE. 481 00:24:28,640 --> 00:24:31,280 THAT CLOSES OFF THE CENTRAL 482 00:24:31,280 --> 00:24:33,840 CHAMBER AND AS I'LL SHOW YOU 483 00:24:33,840 --> 00:24:35,640 THIS OPENS AND CLOSES IN 484 00:24:35,640 --> 00:24:38,280 RESPONSE TO THE BINDING. 485 00:24:38,280 --> 00:24:42,560 NOW AS I MENTIONED -- ABOUT 10% 486 00:24:42,560 --> 00:24:44,640 -- OF THE PROTEINS IT HAS BEEN 487 00:24:44,640 --> 00:24:47,280 LINKED TO A NUMBER OF DISEASE 488 00:24:47,280 --> 00:24:47,960 PARADIGMS. 489 00:24:47,960 --> 00:24:52,160 FOR INSTANCE OUR LAB SHOWS THAT 490 00:24:52,160 --> 00:24:55,680 IT SUPPRESSES TOXICITY AND OTHER 491 00:24:55,680 --> 00:24:57,880 LABS HAVE SHOWN THAT IT 492 00:24:57,880 --> 00:24:59,400 SUPPRESSES AGGREGATION AND 493 00:24:59,400 --> 00:24:59,920 TOXICITY. 494 00:24:59,920 --> 00:25:04,440 AND TO ME THE MOST INTERESTING 495 00:25:04,440 --> 00:25:10,240 ASPECT OF TRIC IS THAT THERE IS 496 00:25:10,240 --> 00:25:13,680 A SUBSET OF PROTEINS THAT CAN 497 00:25:13,680 --> 00:25:16,720 ONLY FOLD WITH TRIC. 498 00:25:16,720 --> 00:25:18,800 THEY COMBINE TO MANY DIFFERENT 499 00:25:18,800 --> 00:25:22,240 TYPES OF CHAPERONES BUT ONLY 500 00:25:22,240 --> 00:25:25,560 TRIC CAN TAKE THEM TO THE NATIVE 501 00:25:25,560 --> 00:25:35,680 STATE. 502 00:25:36,560 --> 00:25:39,160 TRYING TO UNDERSTAND THESE 503 00:25:39,160 --> 00:25:42,120 CHAPERONINS. 504 00:25:42,120 --> 00:25:52,080 WE USE -- -- -- THEY ARE VERY 505 00:25:52,080 --> 00:25:53,520 SIMILAR. 506 00:25:53,520 --> 00:25:58,600 -- -- EACH SUB UNIT HAS THREE 507 00:25:58,600 --> 00:25:59,160 DOMAINS. 508 00:25:59,160 --> 00:26:01,000 ONE BINDS APT. 509 00:26:01,000 --> 00:26:03,760 ONE COMMUNICATES CHANGES. 510 00:26:03,760 --> 00:26:07,160 THIS IS THE LEAD SEGMENT AND 511 00:26:07,160 --> 00:26:08,960 ACTUALLY THERE ARE THESE -- 512 00:26:08,960 --> 00:26:11,280 LITTLE TAILS THAT HAVE NOT BEEN 513 00:26:11,280 --> 00:26:12,960 STRUCTURALLY RESOLVED BUT TURN 514 00:26:12,960 --> 00:26:14,440 OUT TO BE QUITE IMPORTANT AND 515 00:26:14,440 --> 00:26:15,400 I'LL COME BACK TO THAT. 516 00:26:15,400 --> 00:26:19,680 OVER THE YEARS IN COLLABORATIONS 517 00:26:19,680 --> 00:26:25,920 -- -- WE HAVE FOUND STRUCTURES 518 00:26:25,920 --> 00:26:29,440 OF THE OPEN -- STATE AND OF THE 519 00:26:29,440 --> 00:26:31,760 CLOSED STATE AND YOU CAN SEE 520 00:26:31,760 --> 00:26:33,400 THAT THE LEAD IS OPEN IN THE 521 00:26:33,400 --> 00:26:35,160 ABSENCE OF APT. 522 00:26:35,160 --> 00:26:39,800 A FANCY MOVIE OF HOW ATP BINDING 523 00:26:39,800 --> 00:26:43,240 AND HYDROLYSIS BINDS CLOSER OF 524 00:26:43,240 --> 00:26:45,760 THE LEAD WHICH WE'RE TRYING TO 525 00:26:45,760 --> 00:26:48,840 UNDERSTAND BIOCHEMICALLY. 526 00:26:48,840 --> 00:26:52,040 I WANT NOW -- I TOLD YOU THE 527 00:26:52,040 --> 00:26:56,000 REASON -- THEY ARE SIMILAR BUT 528 00:26:56,000 --> 00:27:00,080 IT CANNOT FOLD IN THE OF THE SUB 529 00:27:00,080 --> 00:27:01,600 STRAITS OF TRIC. 530 00:27:01,600 --> 00:27:04,240 WHAT IS SO SPECIAL ABOUT TRIC. 531 00:27:04,240 --> 00:27:05,920 OVER THE YEARS WHAT WE'VE 532 00:27:05,920 --> 00:27:08,920 LEARNED IS THAT THIS -- NATURE 533 00:27:08,920 --> 00:27:15,040 OF TRIC IS EXPLOITED TO GENERATE 534 00:27:15,040 --> 00:27:17,960 ASYMMETRY -- SO AS TO INFLUENCE 535 00:27:17,960 --> 00:27:19,960 THE FOLDING PATHWAY OF THE STUB 536 00:27:19,960 --> 00:27:21,600 STRAIGHT. 537 00:27:21,600 --> 00:27:26,240 SO THE * EARLIEST EXPERIMENTS 538 00:27:26,240 --> 00:27:34,920 THAT LED US TO THESE -- AND WHAT 539 00:27:34,920 --> 00:27:38,360 THEY SHOWED IS THAT OF THE 8 -- 540 00:27:38,360 --> 00:27:41,680 AS SIMILAR AS THEY ARE THERE ARE 541 00:27:41,680 --> 00:27:46,280 FOUR THAT BIND ADP AND FOUR THAT 542 00:27:46,280 --> 00:27:48,720 BIND ADP WITH LOWER. 543 00:27:48,720 --> 00:27:50,720 WHEN THE LEAD CLOSES IT DOESN'T 544 00:27:50,720 --> 00:27:53,080 CLOSE IN A CONCERTED MANNER BUT 545 00:27:53,080 --> 00:27:55,440 MAY CLOSE IN A SEQUENTIAL 546 00:27:55,440 --> 00:27:56,720 MANNER. 547 00:27:56,720 --> 00:27:59,360 ALSO FOUND AND IDENTIFIED THE 548 00:27:59,360 --> 00:28:02,680 BINDING SITE -- AND THEN SHOWED 549 00:28:02,680 --> 00:28:06,200 THAT THE BINDING SITES IN THE 550 00:28:06,200 --> 00:28:09,560 DIFFERENT SUB UNITS HAVE VERY 551 00:28:09,560 --> 00:28:10,800 DIFFERENT PROPERTIES SUGGESTING 552 00:28:10,800 --> 00:28:15,600 THAT THE TRIC CAN BIND IN A 553 00:28:15,600 --> 00:28:19,720 MANNER TO DIFFERENT REGIONS. 554 00:28:19,720 --> 00:28:24,240 AND FINALLY WHEN WE SOLVE THE 555 00:28:24,240 --> 00:28:27,200 ARRANGEMENT OF TRIC WHICH AT THE 556 00:28:27,200 --> 00:28:29,600 TIME WE DIDN'T KNOW -- AND WE 557 00:28:29,600 --> 00:28:34,560 DID THIS IN COLLABORATION -- THE 558 00:28:34,560 --> 00:28:39,320 BOVINE ONE AND WE FOUND THAT 559 00:28:39,320 --> 00:28:42,400 INSIDE OF THE CHAMBER THERE WAS 560 00:28:42,400 --> 00:28:45,320 ALSO ASYMMETRY OF CHARGES WITH 561 00:28:45,320 --> 00:28:48,840 ONE HEMISPHERE BEING POSITIVELY 562 00:28:48,840 --> 00:28:51,160 CHARGED AND ONE BEING NEGATIVELY 563 00:28:51,160 --> 00:28:52,240 CHARGED. 564 00:28:52,240 --> 00:28:54,000 SO THIS STARTED TO MAKE US THINK 565 00:28:54,000 --> 00:29:03,280 THAT PERHAPS FOLDING -- VIA TR 566 00:29:03,280 --> 00:29:08,480 IC WAS NOT THE SAME. 567 00:29:08,480 --> 00:29:10,840 IT TUMBLES AROUND UNTIL IT FOLDS 568 00:29:10,840 --> 00:29:12,720 AND WHEN THE LEAD POPS UP IT 569 00:29:12,720 --> 00:29:14,640 COMES OUT BUT IN REALITY AND 570 00:29:14,640 --> 00:29:16,960 THIS CAME ALSO FROM WORK WITH 571 00:29:16,960 --> 00:29:18,920 NICK DOUGLAS WHEN HE WAS A 572 00:29:18,920 --> 00:29:20,960 STUDENT IN THE LAB THE IDEA WAS 573 00:29:20,960 --> 00:29:23,440 PERHAPS THERE WAS A SUB STRAIGHT 574 00:29:23,440 --> 00:29:27,960 BINDING THAT * THAT TOGETHER 575 00:29:27,960 --> 00:29:30,440 WITH THE CYCLE AND THE UNIQUE 576 00:29:30,440 --> 00:29:33,280 MODE OF RELEASE WHERE ONE SUB 577 00:29:33,280 --> 00:29:36,480 UNIT RELEASED FROM ITS NEIGHBOR 578 00:29:36,480 --> 00:29:38,080 WILL DIRECT THE REMODELING OF 579 00:29:38,080 --> 00:29:41,720 THE POLLY PEPTIDE TO THE SUB 580 00:29:41,720 --> 00:29:45,760 STRAIGHT ALONG THE * SPECIFIC 581 00:29:45,760 --> 00:29:48,600 FOLDING TRAJECTORY. 582 00:29:48,600 --> 00:29:50,160 GUIDED TYPE OF FOLDING. 583 00:29:50,160 --> 00:29:51,800 AND I'M GOING TO TELL YOU A 584 00:29:51,800 --> 00:29:53,160 COUPLE OF RESULTS FROM A PAPER 585 00:29:53,160 --> 00:29:55,440 THAT IS NOT YET PUBLISHED. 586 00:29:55,440 --> 00:29:57,680 MAYBE IT WILL BE PUBLISHED SOME 587 00:29:57,680 --> 00:29:59,760 TIME THIS CENTURY. 588 00:29:59,760 --> 00:30:01,080 THAT ACTUALLY SUGGEST THAT THIS 589 00:30:01,080 --> 00:30:05,080 IS THE CASE HOW TRIC PROMOTES 590 00:30:05,080 --> 00:30:06,440 SUB STRAIGHT FOLDING AND THIS 591 00:30:06,440 --> 00:30:11,520 WAS THE WORK OF -- POSTDOC AND 592 00:30:11,520 --> 00:30:18,600 HAUN ROLE A BIOCHEMIST -- EXPERT 593 00:30:18,600 --> 00:30:21,600 HAHN THAT HAS HIS OWN LAB IN 594 00:30:21,600 --> 00:30:23,760 KOREA. 595 00:30:23,760 --> 00:30:25,800 PURIFIED TRIC. 596 00:30:25,800 --> 00:30:28,600 AND STARTED WITH ONE OF THE 597 00:30:28,600 --> 00:30:29,160 CO-FACTORS. 598 00:30:29,160 --> 00:30:32,000 HE RECONSTITUTED THE FOLDING 599 00:30:32,000 --> 00:30:34,360 PATHWAY INVITRO AND WHAT I'M 600 00:30:34,360 --> 00:30:40,080 SHOWING YOU HERE IS THAT WE 601 00:30:40,080 --> 00:30:41,760 RECONSTITUTED TO LOOK AT THE 602 00:30:41,760 --> 00:30:43,800 STATE OF THE SUBSTRATE DURING 603 00:30:43,800 --> 00:30:48,280 THE FOLDING PATHWAY. 604 00:30:48,280 --> 00:30:49,960 THE SUBSTRATE PROGRESSES FROM A 605 00:30:49,960 --> 00:30:54,360 FULLY UNSTRUCTURED CONFIRMATION 606 00:30:54,360 --> 00:30:56,480 TO A MORE STRUCTURED 607 00:30:56,480 --> 00:30:58,800 CONFIRMATION THAT IS COMPACT BUT 608 00:30:58,800 --> 00:31:05,960 NOT FOLDED WHEN BOUND TO TRIC. 609 00:31:05,960 --> 00:31:06,280 TRICK. 610 00:31:14,000 --> 00:31:17,240 I'M GOING TO GIVE YOU A TIDBIT 611 00:31:17,240 --> 00:31:18,920 OF WHAT WE FOUND BECAUSE IT'S 612 00:31:18,920 --> 00:31:20,080 VERY EXCITING. 613 00:31:20,080 --> 00:31:23,440 IN THE OPEN STATE WE SEE THE 614 00:31:23,440 --> 00:31:25,160 SUBSTRATE OR THE DENSITY SEEMS 615 00:31:25,160 --> 00:31:28,200 TO BE INTERACTING IN THE MIDDLE 616 00:31:28,200 --> 00:31:30,160 OF THE CHAMBER. 617 00:31:30,160 --> 00:31:32,720 THESE ARE NEVER RESOLVED. 618 00:31:32,720 --> 00:31:34,280 THE SUBSTRATE MOVES TO ONE OF 619 00:31:34,280 --> 00:31:35,360 THE CHAMBERS. 620 00:31:35,360 --> 00:31:38,240 WE COULDN'T REALLY RESOLVE THE 621 00:31:38,240 --> 00:31:42,680 SUBSTRATE VERY WELL UNTIL YANG 622 00:31:42,680 --> 00:31:46,120 YANG CHANGE -- COULD IDENTIFY 623 00:31:46,120 --> 00:31:56,520 FOUR DISTINCT FOLDING 624 00:31:59,320 --> 00:31:59,720 INTERMEDIATE. 625 00:31:59,720 --> 00:32:04,440 YOU SEE ONE OF THE DOMAINS 626 00:32:04,440 --> 00:32:04,680 FORMED. 627 00:32:04,680 --> 00:32:07,560 ALL ARE INTERACTING WITH THAT 628 00:32:07,560 --> 00:32:09,600 POSITIVELY CHARGED SITE OF THE 629 00:32:09,600 --> 00:32:11,680 CLOSED CHAMBER THAT I MENTIONED. 630 00:32:11,680 --> 00:32:14,480 THEN WE SEE ANOTHER DOMAIN 631 00:32:14,480 --> 00:32:17,240 FORMED IN INTERMEDIATE TWO AND 632 00:32:17,240 --> 00:32:19,560 ANOTHER IN THREE AND FULLY 633 00:32:19,560 --> 00:32:21,080 FOLDED YOU CAN SUPER IMPOSE IT 634 00:32:21,080 --> 00:32:24,240 WITH A CRYSTAL IN INTERMEDIATE 635 00:32:24,240 --> 00:32:25,880 FOUR. 636 00:32:25,880 --> 00:32:28,640 NOW TRIC DOES NOT FOLD IT BY 637 00:32:28,640 --> 00:32:32,040 RELEASING IN IT THE CHAMBER. 638 00:32:32,040 --> 00:32:35,000 BY HOLDING ON TO EACH ONE OF THE 639 00:32:35,000 --> 00:32:37,480 DOMAINS AND ORGANIZING THIS 640 00:32:37,480 --> 00:32:38,880 STRUCTURE OF THE PROTEIN. 641 00:32:38,880 --> 00:32:42,120 SO HERE IN STATE ONE THE IN 642 00:32:42,120 --> 00:32:45,600 TERMING OF DOMAIN IS FOLDED AND 643 00:32:45,600 --> 00:32:47,760 INTERACTING THROUGH THE 644 00:32:47,760 --> 00:32:48,720 INTERACTIONS. 645 00:32:48,720 --> 00:32:52,160 THE POSITIVE WALL AND THEN THE 646 00:32:52,160 --> 00:32:55,160 -- -- WHICH IS NEGATIVELY 647 00:32:55,160 --> 00:32:57,000 CHARGED IS ALSO INTERACTING WITH 648 00:32:57,000 --> 00:33:03,280 THE POSITIVITILY INTERACTING. 649 00:33:03,280 --> 00:33:05,480 ORGANIZING THE TOP ALLERGY OF 650 00:33:05,480 --> 00:33:08,640 THE * OF THE SUBSTRATE. 651 00:33:08,640 --> 00:33:12,960 AND TO THE VERY TIP OF THE -- -- 652 00:33:12,960 --> 00:33:16,160 IN THE NEXT INTERMEDIATE THE -- 653 00:33:16,160 --> 00:33:19,800 HAS FOLDED AND IT ALSO 654 00:33:19,800 --> 00:33:21,680 INTERRUPTS WITH THE POSITIVELY 655 00:33:21,680 --> 00:33:24,440 CHARGED SIDE OF THE WALL. 656 00:33:24,440 --> 00:33:30,800 SO NOW THE TOPOLOGY -- THIS 657 00:33:30,800 --> 00:33:35,520 LOOKS LIKE NATIVE AND THE MIDDLE 658 00:33:35,520 --> 00:33:38,120 OF THE PROTEIN WE CANNOT SEE IT. 659 00:33:38,120 --> 00:33:41,200 WE KNOW FROM CROSS LINKING IT'S 660 00:33:41,200 --> 00:33:49,400 INTERESIT'SINTERACTING. 661 00:33:49,400 --> 00:33:52,280 SO IN STATE THREE YOU SEE THIS 662 00:33:52,280 --> 00:33:54,400 LITTLE CORE IS BECOMING FORMED 663 00:33:54,400 --> 00:33:59,600 AND NOW WE SEE GTB BINDING TO 664 00:33:59,600 --> 00:34:03,200 THIS NASENT CONFORMER. 665 00:34:03,200 --> 00:34:03,720 *. 666 00:34:03,720 --> 00:34:06,280 SO I THINK THIS IS VERY EXCITING 667 00:34:06,280 --> 00:34:09,720 BECAUSE IT SUGGEST THAT TUBE 668 00:34:09,720 --> 00:34:11,720 LEAN TAR STAR THAT IS A PROTEIN 669 00:34:11,720 --> 00:34:13,240 THAT CAN ONLY FOLD WITH THE 670 00:34:13,240 --> 00:34:15,960 ASSISTANCE OF TRIC IS NOT REALLY 671 00:34:15,960 --> 00:34:16,400 FOLDING. 672 00:34:16,400 --> 00:34:19,560 IT'S FOLLOWING THE DYNAMIC 673 00:34:19,560 --> 00:34:20,120 POTENTIAL. 674 00:34:20,120 --> 00:34:21,720 WHAT WE SPECULATE ALTHOUGH WE 675 00:34:21,720 --> 00:34:23,480 DON'T KNOW BECAUSE WE HAVE NOT 676 00:34:23,480 --> 00:34:25,480 DONE THE BIOPHYSICAL 677 00:34:25,480 --> 00:34:26,960 MEASUREMENTS IS PERHAPS THE 678 00:34:26,960 --> 00:34:28,600 NATIVE STATE IS NOT ONE OF THE 679 00:34:28,600 --> 00:34:29,520 MOST STABLE STATES. 680 00:34:29,520 --> 00:34:32,160 AND WHAT TRIC IS DOING IS BY 681 00:34:32,160 --> 00:34:34,640 THIS SPECIFIC INTERACTION WITH 682 00:34:34,640 --> 00:34:36,880 THE CHAMBER IT'S ACTUALLY 683 00:34:36,880 --> 00:34:40,080 DIRECTING THE DIFFERENT FOLDING 684 00:34:40,080 --> 00:34:42,280 INTERMEDIATES ALONG THE FOLDING 685 00:34:42,280 --> 00:34:42,960 PATHWAY. 686 00:34:42,960 --> 00:34:44,960 ONE THING THAT IS INTERESTING 687 00:34:44,960 --> 00:34:51,400 THERE IS BACTERIA AND IT HAD 688 00:34:51,400 --> 00:34:52,960 LONG BEEN RECOGNIZED THAT THE 689 00:34:52,960 --> 00:34:56,440 DIFFERENCE BETWEEN -- BACTERIA 690 00:34:56,440 --> 00:35:00,000 IS WHERE ALL OF THESE INSERTIONS 691 00:35:00,000 --> 00:35:04,640 ON THE SURFACE OF THE PROTEIN. 692 00:35:04,640 --> 00:35:06,240 THEY INTERACT WITH THE WALL OF 693 00:35:06,240 --> 00:35:11,840 THE CHAMBER THAT LEADS -- THEM 694 00:35:11,840 --> 00:35:12,240 TO FALL. 695 00:35:12,240 --> 00:35:15,520 THIS IS VERY EXCITING TO THINK 696 00:35:15,520 --> 00:35:17,720 ABOUT HOW SOME PROTEINS EVOLVE 697 00:35:17,720 --> 00:35:22,960 AND PERHAPS THE NEW FUNCTIONS OF 698 00:35:22,960 --> 00:35:25,960 TUBE LEAN * AND IT CO-EVOLVED 699 00:35:25,960 --> 00:35:27,960 WITH TRIC TO BE ABLE TO FOLD 700 00:35:27,960 --> 00:35:31,400 WHILE ACQUIRING THESE NEW 701 00:35:31,400 --> 00:35:32,960 FUNCTIONS AND THIS IS A 702 00:35:32,960 --> 00:35:34,160 FASCINATING TOPIC. 703 00:35:34,160 --> 00:35:37,800 I KNOW PATRICK DOLAN LINES TO 704 00:35:37,800 --> 00:35:40,440 THINK ABOUT PROTEIN EVOLUTION. 705 00:35:40,440 --> 00:35:43,080 AND IT IS VERY EXCITING AND KIND 706 00:35:43,080 --> 00:35:46,320 OF LIKE WILL OPEN UP TO THINKING 707 00:35:46,320 --> 00:35:51,320 ABOUT PROTEIN EVOLUTION. 708 00:35:51,320 --> 00:35:53,200 TO FINISH I WANT TO TELL YOU 709 00:35:53,200 --> 00:35:54,320 ABOUT WHAT ARE THE QUESTIONS 710 00:35:54,320 --> 00:35:55,440 THAT WE'RE INTERESTED IN. 711 00:35:55,440 --> 00:35:58,360 WE WANT TO UNDERSTAND HOW TRIC 712 00:35:58,360 --> 00:36:04,560 WORKS FOR OTHER SUBSTRATES. 713 00:36:04,560 --> 00:36:08,080 THERE ARE SOME THAT HAVE THREE 714 00:36:08,080 --> 00:36:10,960 AND FOUR SUB UNITS OF 715 00:36:10,960 --> 00:36:12,160 CHAPERONINS. 716 00:36:12,160 --> 00:36:13,880 HOW TRIC IS REGULATED. 717 00:36:13,880 --> 00:36:18,000 IT'S REDUCED DURING AGING. 718 00:36:18,000 --> 00:36:19,760 SO I THINK THERE COULD BE A LOT 719 00:36:19,760 --> 00:36:21,200 OF INTERESTING THINGS TO DO 720 00:36:21,200 --> 00:36:21,960 THESE. 721 00:36:21,960 --> 00:36:26,160 AND WE RECENTLY COMPLETED A 722 00:36:26,160 --> 00:36:32,080 SMALL MOLECULE REGULATOR SCREEN. 723 00:36:32,080 --> 00:36:35,080 I DIDN'T MENTION THAT TRIC IS 724 00:36:35,080 --> 00:36:44,800 STRONGLY -- REGULATED. 725 00:36:44,800 --> 00:36:47,440 NOW WE WANT TO SWITCH TO WHAT 726 00:36:47,440 --> 00:36:48,880 HAPPENS WHEN PROTEINS CANNOT 727 00:36:48,880 --> 00:36:49,760 FOLD? 728 00:36:49,760 --> 00:36:52,320 I'VE TOLD YOU ABOUT PRODUCTIVE 729 00:36:52,320 --> 00:36:54,280 FOLDING. 730 00:36:54,280 --> 00:36:57,080 WHAT HAPPENS IF A PROTEIN CANNOT 731 00:36:57,080 --> 00:37:00,720 FOLD FOR WHATEVER REASON. 732 00:37:00,720 --> 00:37:02,920 IT CAN EITHER THE SYSTEM CAN 733 00:37:02,920 --> 00:37:06,440 EITHER TARGET IT OR OTHER FORMS 734 00:37:06,440 --> 00:37:09,440 OF DEGRADATION OR KEEP TRYING TO 735 00:37:09,440 --> 00:37:10,200 REFOLD. 736 00:37:10,200 --> 00:37:12,200 SO WE STARTED TO LOOK AT THESE 737 00:37:12,200 --> 00:37:13,800 ALMOST 20 YEARS AGO AND THE 738 00:37:13,800 --> 00:37:16,400 FIRST PERSON WAS AMY 739 00:37:16,400 --> 00:37:18,040 McCLELLAND WHO LOOKED AT A 740 00:37:18,040 --> 00:37:20,880 COUPLE OF MODELS PROTEINS FOR 741 00:37:20,880 --> 00:37:23,360 MISS FOLDING AND THESE ARE ALL 742 00:37:23,360 --> 00:37:25,400 EITHER TEMPERATURE SENSITIVE 743 00:37:25,400 --> 00:37:29,680 PROTEIN OR MU TACT -- OR AN A 744 00:37:29,680 --> 00:37:32,120 SIM BUILD MUTATED TUMOR SUPPRESS 745 00:37:32,120 --> 00:37:32,520 OR. 746 00:37:32,520 --> 00:37:35,160 WHEN THEY MISS FOLD DON'T 747 00:37:35,160 --> 00:37:35,720 AGGREGATE. 748 00:37:35,720 --> 00:37:37,440 THEY JUST GET DEGRADED. 749 00:37:37,440 --> 00:37:39,120 WE WERE ALSO CURIOUS ABOUT 750 00:37:39,120 --> 00:37:41,800 LOOKING AT PROTEINS THAT WHEN 751 00:37:41,800 --> 00:37:44,120 THEY MISS FOLD ACTUALLY 752 00:37:44,120 --> 00:37:51,320 ACCUMULATE AND AGGREGATE. 753 00:37:51,320 --> 00:37:57,080 * THE PROTEINS LOOK VERY 754 00:37:57,080 --> 00:38:02,800 DIFFERENT. 755 00:38:02,800 --> 00:38:05,680 SO WHEN AMY STARTED TO LOOK AT 756 00:38:05,680 --> 00:38:08,200 THIS SHE STARTED WITH THE TUMOR 757 00:38:08,200 --> 00:38:10,440 SUPPRESS OR AND WE KNEW THIS WAS 758 00:38:10,440 --> 00:38:14,160 A PROTEIN THAT TO FOLD NEEDED 759 00:38:14,160 --> 00:38:15,400 TRIC AND THAT IS HOW WE STARTED 760 00:38:15,400 --> 00:38:24,840 TO LOOK AT THIS. 761 00:38:24,840 --> 00:38:27,960 THEY ALSO NEEDED CHAPERONINS TO 762 00:38:27,960 --> 00:38:38,200 GET DEGRADED. 763 00:38:44,040 --> 00:38:47,560 AND THERE SEEMS TO BE A 764 00:38:47,560 --> 00:38:49,040 DEGRADATION CHAPERONE NETWORK 765 00:38:49,040 --> 00:38:50,920 THAT WAS IMPORTANT. 766 00:38:50,920 --> 00:38:52,480 AND OF COURSE THIS WAS 767 00:38:52,480 --> 00:38:55,000 INTERESTING ALSO FROM THE POINT 768 00:38:55,000 --> 00:38:59,240 OF VIEW -- COULD YOU 769 00:38:59,240 --> 00:39:00,880 REFRACTIONAL EYES SOME OF THE 770 00:39:00,880 --> 00:39:02,960 MUTATIONS. 771 00:39:02,960 --> 00:39:05,720 * FAST-FORWARD ANOTHER POSTDOC 772 00:39:05,720 --> 00:39:08,520 IN THE LAB FIGURED OUT WHO WAS 773 00:39:08,520 --> 00:39:17,160 AT THE OO SITE. 774 00:39:17,160 --> 00:39:20,600 HE IDENTIFIED NOT ONLY THAT THIS 775 00:39:20,600 --> 00:39:26,360 CHAPERONES THAT AMY SAW REALLY 776 00:39:26,360 --> 00:39:29,840 PARTICIPATING -- AND ALSO 777 00:39:29,840 --> 00:39:32,640 IDENTIFIED TWO SETS OF -- THAT 778 00:39:32,640 --> 00:39:37,400 NEED TO PRODUCE MIXED LINK 779 00:39:37,400 --> 00:39:39,720 UBIQUITOUS NATION FOR PURPOSES 780 00:39:39,720 --> 00:39:43,200 OF DEGRADATION. 781 00:39:43,200 --> 00:39:46,120 INTERESTING -- THEN GENERATED 782 00:39:46,120 --> 00:39:48,320 NUCLEAR MISS FOLDED PROTEINS AND 783 00:39:48,320 --> 00:39:52,560 IDENTIFIED THE DIFFERENT 784 00:39:52,560 --> 00:39:55,960 CHAPERONE REQUIREMENT AND A 785 00:39:55,960 --> 00:40:00,480 DIFFERENT -- L IgA SE THAT DID 786 00:40:00,480 --> 00:40:05,240 NOT REQUIRE THE LINK BUT 787 00:40:05,240 --> 00:40:09,640 REQUIRED -- TO GET THE PROTEIN 788 00:40:09,640 --> 00:40:12,920 TO THE CYTOPLASM. 789 00:40:12,920 --> 00:40:15,680 THIS IS A SIMPLER SYSTEM. 790 00:40:15,680 --> 00:40:17,720 THE COMPLEXITY OF THIS NETWORK 791 00:40:17,720 --> 00:40:20,440 AND THE THE INTRICACY OF THE 792 00:40:20,440 --> 00:40:22,400 DIFFERENT FACTORS THAT MAKE 793 00:40:22,400 --> 00:40:24,160 DECISIONS ABOUT PROTEIN FATE. 794 00:40:24,160 --> 00:40:27,280 NOW ALL OF THIS EXPERIMENTS YOU 795 00:40:27,280 --> 00:40:29,520 CAN IMAGINE THEM AS HAPPENING IN 796 00:40:29,520 --> 00:40:32,720 THE TESTUD BUT WHEN WE STARTED 797 00:40:32,720 --> 00:40:36,000 LOOK AT IMAGING WHAT HAPPENS TO 798 00:40:36,000 --> 00:40:38,640 MISS FOLDED PROTEINS. 799 00:40:38,640 --> 00:40:41,840 AND THE FIRST PERSON TO DO THIS 800 00:40:41,840 --> 00:40:48,360 IN MY LAB WAS -- LET ME SEE IF I 801 00:40:48,360 --> 00:40:50,720 CAN PLAY THIS MOVIE. 802 00:40:50,720 --> 00:40:51,720 SORRY. 803 00:40:51,720 --> 00:40:52,240 TECHNICAL. 804 00:40:52,240 --> 00:40:53,200 THERE IT IS. 805 00:40:53,200 --> 00:40:55,280 OKAY. 806 00:40:55,280 --> 00:40:57,160 SO THESE ARE TWO DIFFERENT 807 00:40:57,160 --> 00:41:04,560 TEMPERATURE SENSITIVE PROTEINS. 808 00:41:04,560 --> 00:41:07,000 THEY ARE BOTH FOLDED AT THE 809 00:41:07,000 --> 00:41:08,480 BEGINNING OF THE EXPERIMENT AND 810 00:41:08,480 --> 00:41:11,240 THEY ARE DIFFUSED AND FOLDED AND 811 00:41:11,240 --> 00:41:13,760 WHEN THE MOVIE STARTS WE'RE 812 00:41:13,760 --> 00:41:16,720 UNFOLDING THEM BY SHIFTING. 813 00:41:16,720 --> 00:41:20,800 AND THERE'S NO -- PROTEIN 814 00:41:20,800 --> 00:41:22,960 INHIBITION HERE. 815 00:41:22,960 --> 00:41:27,960 YOU CAN SEE THAT -- THEY ARE 816 00:41:27,960 --> 00:41:29,720 ALREADY -- THESE ARE FORMING AND 817 00:41:29,720 --> 00:41:32,520 THE TWO DIFFERENT MISS FOLDED 818 00:41:32,520 --> 00:41:35,240 PROTEINS ARE SEQUESTERED. 819 00:41:35,240 --> 00:41:36,800 THESE ARE NOT AGGREGATES. 820 00:41:36,800 --> 00:41:39,320 THE PROTEIN IS SOLUBLE AND BOUND 821 00:41:39,320 --> 00:41:41,000 TO CHAPERONES. 822 00:41:41,000 --> 00:41:43,000 BUT THIS DOESN'T HAPPEN WITH THE 823 00:41:43,000 --> 00:41:46,520 PROTEIN FLOATING AROUND IN THE 824 00:41:46,520 --> 00:41:47,200 CYTOPLASM. 825 00:41:47,200 --> 00:41:49,840 IT'S SPATIALLY RESTRICT TO 826 00:41:49,840 --> 00:41:55,040 SPECIFIC STRUCTURES IN THE 827 00:41:55,040 --> 00:42:01,480 CYTOPLASM. 828 00:42:01,480 --> 00:42:06,200 SO OVER THE YEARS WORK FROM DON 829 00:42:06,200 --> 00:42:09,280 AND -- HAVE ACTUALLY DEFINED THE 830 00:42:09,280 --> 00:42:13,160 PATHWAY BY WHICH THIS SPATIAL 831 00:42:13,160 --> 00:42:14,000 SEQUESTATION HAPPENS. 832 00:42:14,000 --> 00:42:18,040 WE SEE THAT EVEN IN THE ABSENCE 833 00:42:18,040 --> 00:42:24,080 OF ANY TYPE OF STRESS THIS -- -- 834 00:42:24,080 --> 00:42:27,120 THEY COALESCE AND EVEN AS THEY 835 00:42:27,120 --> 00:42:29,720 GET DEGRADED AND THEY END UP IN 836 00:42:29,720 --> 00:42:34,720 THE VICINITY OF THE NUCLEAR 837 00:42:34,720 --> 00:42:35,360 INVOLVED. 838 00:42:35,360 --> 00:42:37,160 SOLUBLE PROTEINS ARE SEQUESTERED 839 00:42:37,160 --> 00:42:41,000 IN A DIFFERENT INCLUSION. 840 00:42:41,000 --> 00:42:42,880 WHEN THE SYSTEM IS OVERWHELMED 841 00:42:42,880 --> 00:42:47,840 AND WE ACTUALLY EITHER INHIBIT 842 00:42:47,840 --> 00:42:50,400 -- AND THERE IS STRESS GOING ON 843 00:42:50,400 --> 00:42:51,840 THEN THIS LEADS TO A STRUCTURE 844 00:42:51,840 --> 00:42:58,360 CALLED THE JUNK AND RECENTLY -- 845 00:42:58,360 --> 00:43:00,680 ACTUALLY SHOWED THAT THIS JUNKS 846 00:43:00,680 --> 00:43:03,480 NOT ONE BIG INCLUSION BUT A 847 00:43:03,480 --> 00:43:05,760 COLLECTION OF SMALL CUBICLES ALL 848 00:43:05,760 --> 00:43:07,520 COALESCING. 849 00:43:07,520 --> 00:43:09,720 SO BECAUSE OF THIS EXPERIMENT WE 850 00:43:09,720 --> 00:43:12,360 CAN START TO UNDERSTAND WHAT IS 851 00:43:12,360 --> 00:43:15,360 THE FUNCTION OF THESE SPATIAL 852 00:43:15,360 --> 00:43:16,280 SEQUESTATION. 853 00:43:16,280 --> 00:43:20,560 AND THIS WAS WORK DONE BY OUR 854 00:43:20,560 --> 00:43:22,320 POSTDOC IN THE LAB WHO 855 00:43:22,320 --> 00:43:24,120 IDENTIFIED SMALL -- PROTEINS AS 856 00:43:24,120 --> 00:43:28,880 REQUIRED FOR -- FOR -- 857 00:43:28,880 --> 00:43:31,240 FORMATION. 858 00:43:31,240 --> 00:43:37,120 * I'LL PLAY A MOVIE WHERE WE DO 859 00:43:37,120 --> 00:43:42,720 THE SAME EXPERIMENT BUT IN A -- 860 00:43:42,720 --> 00:43:48,520 STRAIN LACKING THE PROTEIN. 861 00:43:48,520 --> 00:43:50,600 YOU WILL SEE THERE IS NO 862 00:43:50,600 --> 00:43:51,280 COALESCENCE. 863 00:43:51,280 --> 00:43:53,960 HOWEVER THE PROTEIN STILL GETS 864 00:43:53,960 --> 00:43:54,360 DEGRADED. 865 00:43:54,360 --> 00:43:57,800 THE CELLS ARE STILL RESISTANT TO 866 00:43:57,800 --> 00:44:00,840 STRESS SO YOU DON'T NEED THIS 867 00:44:00,840 --> 00:44:03,320 FOR DEGRADATION OR STRESS 868 00:44:03,320 --> 00:44:04,080 RESISTANCE. 869 00:44:04,080 --> 00:44:06,560 SO WE WERE WONDERING WHY DO YOU 870 00:44:06,560 --> 00:44:08,440 NEED SPATIAL SEQUESTATION IF YOU 871 00:44:08,440 --> 00:44:10,840 DON'T NEED IT FOR DEGRADATION. 872 00:44:10,840 --> 00:44:12,720 AN EXPERIMENT THAT STEPHANIE DID 873 00:44:12,720 --> 00:44:15,000 IS A COMPETITION EXPERIMENT 874 00:44:15,000 --> 00:44:19,160 WHERE WE MISSED 50% CELLS AND 875 00:44:19,160 --> 00:44:26,200 50% OF MUTANTS -- -- IF WE USE 876 00:44:26,200 --> 00:44:28,200 HSP26 WHICH IS IRRELEVANT TO THE 877 00:44:28,200 --> 00:44:30,800 PROCESS THEN WE GREW THEM FOR 878 00:44:30,800 --> 00:44:33,080 SEVERAL GENERATIONS AND WITH ASK 879 00:44:33,080 --> 00:44:34,680 WHO WINS? 880 00:44:34,680 --> 00:44:36,040 NOBODY WINS. 881 00:44:36,040 --> 00:44:38,480 THE RELATIONSHIP OF MUTANT TO -- 882 00:44:38,480 --> 00:44:39,320 IS MAINTAINED. 883 00:44:39,320 --> 00:44:42,720 WHEN WE DO THIS FOR THE -- 884 00:44:42,720 --> 00:44:44,120 PROTEIN UNDER NO STRESS 885 00:44:44,120 --> 00:44:45,360 CONDITIONS SAME THING. 886 00:44:45,360 --> 00:44:47,600 7 DOESN'T REALLY MATTER BUT WHEN 887 00:44:47,600 --> 00:44:49,840 WE PUT THE CELLS UNDER STRESS 888 00:44:49,840 --> 00:44:53,360 THE CELLS THAT CANNOT FORM THE 889 00:44:53,360 --> 00:44:56,360 SPATIAL SEQUESTATION OF MISS 890 00:44:56,360 --> 00:44:59,200 FOLDED PROTEINS ARE LESS FIT AND 891 00:44:59,200 --> 00:45:01,200 ARE OUTCOMPETED BY THE WILD 892 00:45:01,200 --> 00:45:01,600 TYPE. 893 00:45:01,600 --> 00:45:03,200 SO THE CONCLUSION OF THESE IS 894 00:45:03,200 --> 00:45:08,800 THAT SPECIFIC CHAPERONE PATHWAYS 895 00:45:08,800 --> 00:45:11,880 -- CLEAR DECISIONS IN THE CITE 896 00:45:11,880 --> 00:45:13,880 PLASM AND THEY ARE DIFFERENT. 897 00:45:13,880 --> 00:45:16,280 THAT PROTEIN QUALITY CONTROL IS 898 00:45:16,280 --> 00:45:18,000 SPATIALLY ORGANIZED AND THAT 899 00:45:18,000 --> 00:45:20,400 SPATIAL SEQUESTATION IS A 900 00:45:20,400 --> 00:45:22,680 PROTECTIVE MECHANISM. 901 00:45:22,680 --> 00:45:24,720 AND SO SHOW YOU THIS HAS SOME 902 00:45:24,720 --> 00:45:26,280 RELEVANCE I WANT TO BRING UP 903 00:45:26,280 --> 00:45:33,520 SOME EXPERIMENTS WHERE THEY 904 00:45:33,520 --> 00:45:39,240 COLORADO COLLABORATED -- AND TO 905 00:45:39,240 --> 00:45:42,040 OUR SURPRISE IF YOU DO THESE YOU 906 00:45:42,040 --> 00:45:44,720 CAN SEE A BIG INCLUSION AND THEN 907 00:45:44,720 --> 00:45:48,800 YOU SEE WHAT IS REFERRED TO IS A 908 00:45:48,800 --> 00:45:52,080 SOLUBLE DIFFUSED FLUORESCENCE. 909 00:45:52,080 --> 00:45:54,600 THEY FOUND ACTUALLY THERE WAS NO 910 00:45:54,600 --> 00:45:57,600 SOLUBLE DIFFUSED FLUORESCENCE. 911 00:45:57,600 --> 00:46:01,640 IT IS A SERIES OF SERIES OF 912 00:46:01,640 --> 00:46:02,320 SMALL STRUCTURES. 913 00:46:02,320 --> 00:46:05,960 THEY ARE NOT BIG INCLUSIONS BUT 914 00:46:05,960 --> 00:46:08,160 THEY DO LOOK SMALL IN NATURE. 915 00:46:08,160 --> 00:46:11,120 AND IN COLLABORATION WITH LESLIE 916 00:46:11,120 --> 00:46:13,440 THOMPSON WE HAD DEVELOPED A 917 00:46:13,440 --> 00:46:17,080 CHAPERONE STRATEGY TO RESCUE THE 918 00:46:17,080 --> 00:46:23,120 TOXICITY AND WHEN WE ADDED THESE 919 00:46:23,120 --> 00:46:27,440 EXPRESSING MUTANT HUNTINGTON AND 920 00:46:27,440 --> 00:46:29,000 SUBJECTED THEM WE FOUND THAT 921 00:46:29,000 --> 00:46:31,440 THIS CHAPERONE DOMAIN LONG STORY 922 00:46:31,440 --> 00:46:33,680 AND I CAN TELL YOU MORE BUT THIS 923 00:46:33,680 --> 00:46:37,800 IS A CHAPERONE THAT SUPPRESSES 924 00:46:37,800 --> 00:46:39,800 HUNTINGTON TOXICITY PRIMARILY 925 00:46:39,800 --> 00:46:44,280 REDUCE THE AMOUNT OF THE SOLUBLE 926 00:46:44,280 --> 00:46:46,360 DIFFUSED LITTLE STRUCTURES. 927 00:46:46,360 --> 00:46:49,240 WHICH AGAIN SUGGEST THAT PERHAPS 928 00:46:49,240 --> 00:46:51,280 SEQUESTATION IS A PROTECTIVE 929 00:46:51,280 --> 00:46:51,680 PATHWAY. 930 00:46:51,680 --> 00:46:53,880 SO BASICALLY WE'RE INTERESTED IN 931 00:46:53,880 --> 00:46:56,720 UNDERSTANDING THE PATHWAYS THAT 932 00:46:56,720 --> 00:46:58,200 PROMOTE QUALITY CONTROL 933 00:46:58,200 --> 00:46:59,440 OBVIOUSLY THE BIG QUESTION IS 934 00:46:59,440 --> 00:47:01,880 WHY IS THERE TOXICITY FROM THESE 935 00:47:01,880 --> 00:47:03,960 FOLDED PROTEINS AND WE'RE VERY 936 00:47:03,960 --> 00:47:08,080 INTERESTED IN LOOKING FORWARD 937 00:47:08,080 --> 00:47:09,560 ABOUT THIS STRUCTURE OF THESE 938 00:47:09,560 --> 00:47:09,960 INCLUSIONS. 939 00:47:09,960 --> 00:47:16,120 7 HOW MUCH MORE TIME DO I HAVE? 940 00:47:16,120 --> 00:47:16,600 OKAY. 941 00:47:16,600 --> 00:47:21,360 SO I'M GOING TO GIVE YOU A A 942 00:47:21,360 --> 00:47:25,080 LITTLE -- ALL OF THIS STUFF SO 943 00:47:25,080 --> 00:47:27,360 FAR WE TRY TO UNDERSTAND BASIC 944 00:47:27,360 --> 00:47:31,040 PRINCIPLES BUT AS YOU KNOW -- IT 945 00:47:31,040 --> 00:47:33,280 IS REGULATED IN MANY INTERESTING 946 00:47:33,280 --> 00:47:35,280 WAYS AND THAT COULD BE USEFUL 947 00:47:35,280 --> 00:47:36,760 FOR THERAPEUTICS. 948 00:47:36,760 --> 00:47:38,760 THESE ARE A NUMBER OF THINGS 949 00:47:38,760 --> 00:47:39,800 THAT WE HAVE BEEN INTERESTED IN 950 00:47:39,800 --> 00:47:40,760 THE LAB. 951 00:47:40,760 --> 00:47:42,040 WE'RE VERY INTERESTED IN 952 00:47:42,040 --> 00:47:44,360 UNDERSTANDING AND APPLYING OUR 953 00:47:44,360 --> 00:47:50,080 BASIC UNDERSTANDING TO TOXICITY 954 00:47:50,080 --> 00:47:53,600 DURING NEURODEGENERATION AND WHY 955 00:47:53,600 --> 00:47:57,280 -- IT DECLINES DURING AGING AND 956 00:47:57,280 --> 00:48:02,640 HOW CAN THESE DECLINES SYNERGIZE 957 00:48:02,640 --> 00:48:05,520 AND -- FOR MANY YEARS AND 958 00:48:05,520 --> 00:48:07,720 COLLABORATION TO TRY TO 959 00:48:07,720 --> 00:48:10,520 UNDERSTAND HOW VIRUSES -- AND 960 00:48:10,520 --> 00:48:13,680 CAN WE EXPLOIT THAT FOR 961 00:48:13,680 --> 00:48:14,600 THERAPEUTIC PURPOSES. 962 00:48:14,600 --> 00:48:18,600 7 SO I'LL MENTION A COUPLE OF 963 00:48:18,600 --> 00:48:19,840 THINGS WITHOUT GOING INTO A LOT 964 00:48:19,840 --> 00:48:21,240 OF DETAIL. 965 00:48:21,240 --> 00:48:24,040 ONE OF -- HAS BEEN TRYING TO 966 00:48:24,040 --> 00:48:27,240 UNDERSTAND HOW THE PROCESS OF 967 00:48:27,240 --> 00:48:29,320 TRANSLATION COUPLED FOLDING AND 968 00:48:29,320 --> 00:48:31,080 BIOGEN SIS IS IMPACTED BY 969 00:48:31,080 --> 00:48:36,160 DISEASE AND YOU CAN THINK OF THE 970 00:48:36,160 --> 00:48:41,680 * RIBOSOME AS A SUPER MOLECULAR 971 00:48:41,680 --> 00:48:42,400 PROTEIN MACHINE. 972 00:48:42,400 --> 00:48:46,840 WE HAVE A PAPER SHOWING THAT -- 973 00:48:46,840 --> 00:48:48,760 HUNTINGTON DISEASE PROMOTES 974 00:48:48,760 --> 00:48:52,160 TRANSLATION DISFUNCTION AND 975 00:48:52,160 --> 00:48:55,280 PROMOTES WIDESPREAD RIBOSOME -- 976 00:48:55,280 --> 00:48:57,800 THAT IS TOXIC. 977 00:48:57,800 --> 00:49:02,760 AGING ALSO DISRUPTS -- AND THIS 978 00:49:02,760 --> 00:49:11,440 LEADS TO PROTEIN -- ---AND LAST 979 00:49:11,440 --> 00:49:15,520 YEAR -- SHOWED THAT FOR A NUMBER 980 00:49:15,520 --> 00:49:19,600 OF DIFFERENT VIRUSES THE 981 00:49:19,600 --> 00:49:22,760 INFECTION TOTALLY REMODELS THE 982 00:49:22,760 --> 00:49:23,920 TRANSLATIONAL APPARATUS AND 983 00:49:23,920 --> 00:49:27,880 THERE ARE SOME MODULES SUCH AS 984 00:49:27,880 --> 00:49:29,640 TRANSLATION INITIATION -- 985 00:49:29,640 --> 00:49:33,080 QUALITY CONTROL THAT ARE EVICTED 986 00:49:33,080 --> 00:49:34,440 FROM THE RIBOSOME AND SPECIFIC 987 00:49:34,440 --> 00:49:37,480 CHAPERONES THAT ARE RECRUITED TO 988 00:49:37,480 --> 00:49:38,960 RIBOSOMES DURING VIRAL INFECTION 989 00:49:38,960 --> 00:49:41,840 AND THE INTERESTING ASPECT OF 990 00:49:41,840 --> 00:49:45,520 THE WORK IS THAT THE ENTIRE 991 00:49:45,520 --> 00:49:49,040 COLLAGEN -- MACHINERY IS 992 00:49:49,040 --> 00:49:51,680 HIGHJACKED BY VIRUSES TO HELP 993 00:49:51,680 --> 00:49:53,760 PROMOTE THE -- AND IF YOU 994 00:49:53,760 --> 00:49:56,920 INHIBIT THIS MACHINERY HALF OF 995 00:49:56,920 --> 00:49:59,520 THE VIRAL PROTECT TEEN MISS 996 00:49:59,520 --> 00:50:01,040 FOLDS. 997 00:50:01,040 --> 00:50:05,520 * * I'M GOING TO SKIP THE AGING 998 00:50:05,520 --> 00:50:13,520 PART. 999 00:50:13,520 --> 00:50:17,720 WHEN WE COMPARE -- THE PROTEINS 1000 00:50:17,720 --> 00:50:19,800 THAT WERE PREVIOUSLY IDENTIFIED 1001 00:50:19,800 --> 00:50:23,560 TO AGGREGATE WITH AGING WORMS WE 1002 00:50:23,560 --> 00:50:25,120 SEE HIGH LEVELS OF CORRELATION. 1003 00:50:25,120 --> 00:50:32,400 SO WE'RE NOW HYPOTHESISSING -- 1004 00:50:32,400 --> 00:50:37,760 ONE OF THE REASONS THAT YOU SEE 1005 00:50:37,760 --> 00:50:41,080 ENHANCED DEGRADATION. 1006 00:50:41,080 --> 00:50:43,640 I'LL FINISH WITH SOME TAKE HOME 1007 00:50:43,640 --> 00:50:45,040 MESSAGES. 1008 00:50:45,040 --> 00:50:49,520 I HOPE I CONVINCED YOU THAT THE 1009 00:50:49,520 --> 00:50:51,880 MACHINERY PROFOUNDLY INFLUENCES 1010 00:50:51,880 --> 00:50:53,160 PROTEIN FATE. 1011 00:50:53,160 --> 00:50:55,360 IT KEEPS MUTANT PROTEINS 1012 00:50:55,360 --> 00:50:56,040 FUNCTIONAL. 1013 00:50:56,040 --> 00:50:57,480 IT PREVENTS AGGREGATION. 1014 00:50:57,480 --> 00:51:03,840 THIS IS GOOD IN A YOUNG PROTECT 1015 00:51:03,840 --> 00:51:05,160 YUM. 1016 00:51:05,160 --> 00:51:08,600 * IT CAN BE BAD IN A CANCER 1017 00:51:08,600 --> 00:51:11,280 CELL. 1018 00:51:11,280 --> 00:51:15,280 IF YOU INHIBIT THE CAPACITY AND 1019 00:51:15,280 --> 00:51:17,520 YOU CAN THINK OF THESE AS BEING 1020 00:51:17,520 --> 00:51:19,520 IN AGING WHERE YOU NOW START TO 1021 00:51:19,520 --> 00:51:23,040 SHOW THE PHENOTYPES OF THE 1022 00:51:23,040 --> 00:51:25,000 MUTANTS THAT WE HAVE AND THE 1023 00:51:25,000 --> 00:51:26,520 MACHINERY KEEPS FUNCTIONAL OR 1024 00:51:26,520 --> 00:51:29,080 WHEN YOU INTERVENE WITH DRUGS TO 1025 00:51:29,080 --> 00:51:32,960 TREAT VIRUSES OR CANCER YOU WILL 1026 00:51:32,960 --> 00:51:35,840 AGGREGATE OR CAUSE DEGRADATION 1027 00:51:35,840 --> 00:51:37,480 OF MANY THESE PROTEINS. 1028 00:51:37,480 --> 00:51:44,160 I THINK THIS CAN BE A VERY POW 1029 00:51:44,160 --> 00:51:44,960 POWERFUL SYSTEM. 1030 00:51:44,960 --> 00:51:46,840 WE NEED TO GET PAST THINKING 1031 00:51:46,840 --> 00:51:49,720 THAT THE MUTATION OR THE PROTEIN 1032 00:51:49,720 --> 00:51:51,840 IS JUST A FUNCTION OF ITS OWN 1033 00:51:51,840 --> 00:51:52,400 STABILITY. 1034 00:51:52,400 --> 00:51:54,360 IT'S A COMBINES OF ITS OWN 1035 00:51:54,360 --> 00:51:56,800 STABILITY AND THIS I TOOK FROM A 1036 00:51:56,800 --> 00:51:59,120 REVIEW FROM JEFF KELLY -- AND 1037 00:51:59,120 --> 00:52:03,880 THE CONDITIONS IN THE CELL SO 1038 00:52:03,880 --> 00:52:07,640 THE LEVEL OF CHAPERONES WILL ALL 1039 00:52:07,640 --> 00:52:11,600 KEEP THE PROTEIN WITHIN ITS 1040 00:52:11,600 --> 00:52:17,200 FUNCTIONAL RANGE. 1041 00:52:17,200 --> 00:52:22,960 SO IN THINKING ABOUT THIS -- WE 1042 00:52:22,960 --> 00:52:24,800 CANNOT THINK ABOUT THIS IN 1043 00:52:24,800 --> 00:52:25,760 SIMPLISTIC WAYS. 1044 00:52:25,760 --> 00:52:29,960 WE NEED TO THINK ABOUT THE DECAY 1045 00:52:29,960 --> 00:52:33,040 AS A VICIOUS CIRCLE -- WORKING 1046 00:52:33,040 --> 00:52:34,840 TOGETHER WITH AGING TO INCREASE 1047 00:52:34,840 --> 00:52:37,960 AGGREGATION AND INCREASE PROTEIN 1048 00:52:37,960 --> 00:52:38,960 DYSFUNCTION BECAUSE THERE IS 1049 00:52:38,960 --> 00:52:43,160 THIS -- CAPACITY -- THE VICIOUS 1050 00:52:43,160 --> 00:52:46,240 CIRCLE GOES AHEAD AND MAYBE I'LL 1051 00:52:46,240 --> 00:52:49,200 JUST END BY SAYING THAT IN THIS 1052 00:52:49,200 --> 00:52:52,200 LIGHT -- THE PRESENCE OF 1053 00:52:52,200 --> 00:52:56,200 AGGREGATES IN AN AGE -- DECEASED 1054 00:52:56,200 --> 00:52:57,840 CELL MAY NOT BE THE PROBLEM. 1055 00:52:57,840 --> 00:53:00,360 IT MAY BE AN EMERGING PROPERTY 1056 00:53:00,360 --> 00:53:04,520 OF DYSFUNCTIONAL -- MACHINERY. 1057 00:53:04,520 --> 00:53:06,880 IN A YOUNG CELL YOU HAVE MANY 1058 00:53:06,880 --> 00:53:08,320 OPTIONS WHERE YOU HAVE MISS 1059 00:53:08,320 --> 00:53:11,120 FOLDED PROTEINS AND AS THE CELL 1060 00:53:11,120 --> 00:53:13,880 AGES AND THE CAPACITY GOES DOWN 1061 00:53:13,880 --> 00:53:16,560 THEN THE ONLY FUNCTION IS A 1062 00:53:16,560 --> 00:53:18,640 SEQUESTATION AND YOU MAY BE LEFT 1063 00:53:18,640 --> 00:53:21,400 WITH THIS TYPE OF AGGREGATE. 1064 00:53:21,400 --> 00:53:22,600 WE'RE INTERESTED IN 1065 00:53:22,600 --> 00:53:24,240 UNDERSTANDING THE BASIC ASPECT 1066 00:53:24,240 --> 00:53:25,840 OF THE FUNCTION OF THE MACHINERY 1067 00:53:25,840 --> 00:53:29,800 AND ALSO UNDER HOW IT GOES. 1068 00:53:29,800 --> 00:53:31,360 I'LL ACKNOWLEDGE ALL OF THE 1069 00:53:31,360 --> 00:53:32,960 PEOPLE THAT DID THE WORK. 1070 00:53:32,960 --> 00:53:37,280 I THINK I MENTIONED EVERYBODY. 1071 00:53:37,280 --> 00:53:42,720 THESE ARE THE MAIN CONTRIBUTORS. 1072 00:53:42,720 --> 00:53:45,760 I HAVE TO ACKNOWLEDGE OUR 1073 00:53:45,760 --> 00:53:49,200 COLLABORATORS. 1074 00:53:49,200 --> 00:53:53,520 AND I REALLY NEED TO THAT AND 1075 00:53:53,520 --> 00:53:55,040 NIH FOR FUNDING. 1076 00:53:55,040 --> 00:53:57,480 IT'S PAINFUL BUT WITHOUT IT 1077 00:53:57,480 --> 00:53:59,040 NOTHING OF THIS COULD HAPPEN SO 1078 00:53:59,040 --> 00:54:02,920 I HAVE TO THANK THEM ALL THAT 1079 00:54:02,920 --> 00:54:05,040 GAVE US MONEY TO DO THIS WORK. 1080 00:54:05,040 --> 00:54:07,960 [ APPLAUSE ] 1081 00:54:07,960 --> 00:54:09,600 >> THANK YOU AND ANYONE HERE IS 1082 00:54:09,600 --> 00:54:12,760 FREE TO USE THE MICS. 1083 00:54:12,760 --> 00:54:14,000 WE HAVE SEVERAL QUESTIONS COMING 1084 00:54:14,000 --> 00:54:15,200 IN. 1085 00:54:15,200 --> 00:54:17,240 ONE IS A COMMENT THAT JUST CAME 1086 00:54:17,240 --> 00:54:18,960 NCATS PLEASE INVITE HER BACK TO 1087 00:54:18,960 --> 00:54:20,880 TEACH US MORE ABOUT AGING AND 1088 00:54:20,880 --> 00:54:26,280 THE SLIDES SHE JUST TEASED US 1089 00:54:26,280 --> 00:54:26,720 WITH. 1090 00:54:26,720 --> 00:54:28,720 MADE MY DAY. 1091 00:54:28,720 --> 00:54:31,160 GIVE HER A ROUND OF APPLAUSE. 1092 00:54:31,160 --> 00:54:32,040 >> THIS IS FANTASTIC. 1093 00:54:32,040 --> 00:54:33,040 THANK YOU SO MUCH. 1094 00:54:33,040 --> 00:54:35,240 AND I HOPE I DIDN'T MISS IT BUT 1095 00:54:35,240 --> 00:54:39,000 I WAS STRUCK BY YOUR STRUCTURES 1096 00:54:39,000 --> 00:54:42,080 AND FOLDING -- AND I WAS 1097 00:54:42,080 --> 00:54:43,880 WONDERING 10% OF HUMAN PROTEINS 1098 00:54:43,880 --> 00:54:47,720 ARE USING THE SAME KIND OF 1099 00:54:47,720 --> 00:54:48,720 MECHANISM. 1100 00:54:48,720 --> 00:54:51,320 SO ARE THEY STRUCTURALLY THE 1101 00:54:51,320 --> 00:54:54,280 SAME OR IS THERE STRUCTURAL -- 1102 00:54:54,280 --> 00:54:55,560 IN TRIC. 1103 00:54:55,560 --> 00:54:57,600 CAN YOU COMMENT ON HOW YOU 1104 00:54:57,600 --> 00:54:58,600 ENVISION THAT WORKING? 1105 00:54:58,600 --> 00:55:00,080 >> THAT IS A VERY GOOD QUESTION 1106 00:55:00,080 --> 00:55:05,440 AND A LOT OF THINGS TO THINK B. 1107 00:55:05,440 --> 00:55:07,680 WE HAVE THE STRUCTURE OF TUBE 1108 00:55:07,680 --> 00:55:10,040 LEAN * AND WE HAVE AN ONGOING 1109 00:55:10,040 --> 00:55:13,000 STRUCTURE OF ACTING SO, THAT IS 1110 00:55:13,000 --> 00:55:14,880 THE EXTENT OF MY UNDERSTANDING 1111 00:55:14,880 --> 00:55:18,160 OF FOLDING INTERMEDIATES. 1112 00:55:18,160 --> 00:55:23,680 WE HAD PUBLISHED A PAPER -- SO 1113 00:55:23,680 --> 00:55:28,520 IT -- PROTEIN FROM REAL VIRUSES 1114 00:55:28,520 --> 00:55:31,800 ALSO NEEDS TRIC TO FOLD. 1115 00:55:31,800 --> 00:55:34,360 ACTUALLY BINDS TO DIFFERENT 1116 00:55:34,360 --> 00:55:37,880 REGIONS WITHIN THE CHAMBER. 1117 00:55:37,880 --> 00:55:40,240 MAYBE DIFFERENT PROTEINS THAT 1118 00:55:40,240 --> 00:55:42,840 NEED TRIC TO FOLD WILL INTERACT 1119 00:55:42,840 --> 00:55:45,120 WITH DIFFERENT REGIONS AND MAYBE 1120 00:55:45,120 --> 00:55:50,200 THIS IS WHAT THE -- -- DID IT 1121 00:55:50,200 --> 00:55:52,040 GENERATED THE POSSIBILITY THAT 1122 00:55:52,040 --> 00:55:54,880 DIFFERENT PROTEINS WILL BIND IN 1123 00:55:54,880 --> 00:55:56,200 DIFFERENT REGIONS. 1124 00:55:56,200 --> 00:55:59,600 WITH THAT SAID A LARGE NUMBER OF 1125 00:55:59,600 --> 00:56:02,600 TRIC SUBSTRATES THAT WE SEE 1126 00:56:02,600 --> 00:56:05,400 DIRECTLY -- PROBABLY ALL THEY 1127 00:56:05,400 --> 00:56:11,080 NEED IS -- CHAPERONE. 1128 00:56:11,080 --> 00:56:16,440 SO A LOT OF PROTEINS -- 1129 00:56:16,440 --> 00:56:19,480 ---ENZYMES WHOSE BACTERIAL HOMO 1130 00:56:19,480 --> 00:56:25,840 LOGS -- SO FOR THE EVOLUTION OF 1131 00:56:25,840 --> 00:56:28,480 TRIC IT'S NOT THAT TRICKS 1132 00:56:28,480 --> 00:56:32,160 INVOLVED -- TUBE LEAN * AND 1133 00:56:32,160 --> 00:56:35,920 OTHER SUBSTRATES EVOLVED AS TRIC 1134 00:56:35,920 --> 00:56:38,000 DIVERSITY FIGHTERS -- TO USE 1135 00:56:38,000 --> 00:56:39,600 SPECIFIC FEATURES OF THE CHAMBER 1136 00:56:39,600 --> 00:56:42,280 OF THE SUBSTRATE BINDING SITE OF 1137 00:56:42,280 --> 00:56:43,720 THE TAILS. 1138 00:56:43,720 --> 00:56:46,000 BUT TRIC ALSO MAINTAIN THE 1139 00:56:46,000 --> 00:56:49,440 ABILITY TO BIND AND ENGULF 1140 00:56:49,440 --> 00:56:52,080 CHAPERONES AND WHAT THEY NEED IS 1141 00:56:52,080 --> 00:56:55,280 TO BIND AND FALL TO THE NATIVE 1142 00:56:55,280 --> 00:56:57,280 STATE AND BE FOLDED. 1143 00:56:57,280 --> 00:56:59,360 SO I DON'T THINK EVERY PROTEIN 1144 00:56:59,360 --> 00:57:05,640 THAT INTERACTS IS AN OBLIGATE 1145 00:57:05,640 --> 00:57:15,800 SUBSTRATE. 1146 00:57:20,400 --> 00:57:23,640 WHEN WE LOOK AT HOW THEY BIND 6 1147 00:57:23,640 --> 00:57:26,200 THESE ARE COMPLETELY DIFFERENT 1148 00:57:26,200 --> 00:57:28,640 PROTEINS BY TRIC BINDS IN THE 1149 00:57:28,640 --> 00:57:30,960 SAME PLACE OF THE BARREL OF THE 1150 00:57:30,960 --> 00:57:31,520 BLADES. 1151 00:57:31,520 --> 00:57:34,920 SO I THINK THERE ARE A LOT OF 1152 00:57:34,920 --> 00:57:35,840 VERY COOL QUESTIONS. 1153 00:57:35,840 --> 00:57:37,800 >> THANK YOU. 1154 00:57:37,800 --> 00:57:39,120 >> TWO QUESTIONS ON-LINE. 1155 00:57:39,120 --> 00:57:41,200 BECAUSE IT MIGHT BE RELATED 1156 00:57:41,200 --> 00:57:43,360 TOGETHER IN WHAT HE JUST ASKED. 1157 00:57:43,360 --> 00:57:45,920 FIRST IS WHAT IS THE FOLDING 1158 00:57:45,920 --> 00:57:47,200 COMPLEXITY OF THE CHAPERONE 1159 00:57:47,200 --> 00:57:47,800 PROTEINS? 1160 00:57:47,800 --> 00:57:50,160 DO THEY ALSO REQUIRE CHAPERONS 1161 00:57:50,160 --> 00:57:51,120 FOR FOLDING? 1162 00:57:51,120 --> 00:57:54,240 AND DO ALL-PRO TEENS FOLDED BY 1163 00:57:54,240 --> 00:57:56,000 THE TRIC CHAPERONE SHARE A 1164 00:57:56,000 --> 00:57:57,440 SIMILAR STRUCTURE THAT ALLOWS 1165 00:57:57,440 --> 00:58:00,760 TRIC TO MONITOR FOLDING AND A 1166 00:58:00,760 --> 00:58:03,880 SIMILAR MECHANISM FOR ALL 1167 00:58:03,880 --> 00:58:04,440 SUBSTRATES 1168 00:58:04,440 --> 00:58:06,640 SOME OF THEM AT LEAST ALSO NEED 1169 00:58:06,640 --> 00:58:09,520 CHAPERONES. 1170 00:58:09,520 --> 00:58:14,560 WE KNOW THAT -- 1171 00:58:14,560 --> 00:58:15,160 CO-TRANSLATIONALLY. 1172 00:58:15,160 --> 00:58:17,480 THEY ALSO NEED -- THEY ARE JUST 1173 00:58:17,480 --> 00:58:19,560 PROTEINS LIKE OTHER PROTEINS MAY 1174 00:58:19,560 --> 00:58:22,080 NEED CHAPERONES TO FOLD. 1175 00:58:22,080 --> 00:58:24,040 WITH REGARD TO TRIC YES THERE 1176 00:58:24,040 --> 00:58:26,000 ARE SOME DOMAINS THAT ARE OVER 1177 00:58:26,000 --> 00:58:27,960 REPRESENTED IN TRIC. 1178 00:58:27,960 --> 00:58:32,800 SO THE BETTER PROPELLER -- 40 1179 00:58:32,800 --> 00:58:36,400 PROTEINS -- WHICH TUBE LEAN AND 1180 00:58:36,400 --> 00:58:39,640 OKAY TEEN HAVE -- FOLDS *. 1181 00:58:39,640 --> 00:58:42,840 LOTS OF RNA AND DNA BINDING 1182 00:58:42,840 --> 00:58:46,080 PROTEINS. 1183 00:58:46,080 --> 00:58:48,640 THEY SEEM TO ME TO BE ABLE TO 1184 00:58:48,640 --> 00:58:50,440 FOLD. 1185 00:58:50,440 --> 00:58:53,840 BUT THERE ISN'T ONE TYPE OF 1186 00:58:53,840 --> 00:58:54,400 DOMAIN. 1187 00:58:54,400 --> 00:58:56,200 THERE ARE OTHER PROTEINS THAT 1188 00:58:56,200 --> 00:58:57,040 DON'T NEED TRIC. 1189 00:58:57,040 --> 00:58:59,600 IT'S NOT JUST THE FOLD. 1190 00:58:59,600 --> 00:59:03,440 THERE MUST BE ELEMENTS THAT -- 1191 00:59:03,440 --> 00:59:06,560 INTERFERE WITH THE FOLDING 1192 00:59:06,560 --> 00:59:07,680 PATHWAY. 1193 00:59:07,680 --> 00:59:12,120 THAT MAKE THEM NEED TRIC. 1194 00:59:12,120 --> 00:59:13,520 >> AWESOME. 1195 00:59:13,520 --> 00:59:15,680 MY QUESTION IS ALSO ABOUT TRIC. 1196 00:59:15,680 --> 00:59:18,760 WITH SO MANY DIFFERENT FOLDING 1197 00:59:18,760 --> 00:59:21,320 SUBSTRATES HOW DOES TRIC KNOW 1198 00:59:21,320 --> 00:59:23,400 WHEN EACH PEPTIDE IS DONE 1199 00:59:23,400 --> 00:59:24,520 FOLDING? 1200 00:59:24,520 --> 00:59:25,840 HOW DOES IT SENSE? 1201 00:59:25,840 --> 00:59:27,800 IS IT ON A TIMER? 1202 00:59:27,800 --> 00:59:34,640 DOES A FOLDED PROTEIN -- WHAT IS 1203 00:59:34,640 --> 00:59:36,080 AUGMENTED? 1204 00:59:36,080 --> 00:59:36,760 >> HMMM. 1205 00:59:36,760 --> 00:59:40,960 GOOD QUESTION. 1206 00:59:40,960 --> 00:59:44,880 SO THE CYCLE -- OF TRICKLE POSES 1207 00:59:44,880 --> 00:59:47,080 A TIMER. 1208 00:59:47,080 --> 00:59:51,760 WE CECE THIS -- THIS WOULD BE 1209 00:59:51,760 --> 00:59:54,840 NICER DONE IN VERY VOTE BUT 1210 00:59:54,840 --> 01:00:00,720 WE'VE DONE INVITRO EXPERIMENTS 1211 01:00:00,720 --> 01:00:10,200 THAT SUGGEST -- PROBABLY AFTER 1212 01:00:10,200 --> 01:00:12,840 THAT THEY GET DEGRADED. 1213 01:00:12,840 --> 01:00:15,320 BUT THERE IS SOME PROTEINS THAT 1214 01:00:15,320 --> 01:00:19,000 THEY FOLD AND THEY STAY BOUND TO 1215 01:00:19,000 --> 01:00:19,520 TRIC. 1216 01:00:19,520 --> 01:00:23,520 SO MANY SUBSTRATES ARE -- OF 1217 01:00:23,520 --> 01:00:25,720 LARGE COMPLEXES. 1218 01:00:25,720 --> 01:00:28,360 FOR THOSE THERE MUST BE A 1219 01:00:28,360 --> 01:00:29,960 MECHANISM TO KICK THEM OFF. 1220 01:00:29,960 --> 01:00:32,600 EITHER THE PARTNER COMES IN AND 1221 01:00:32,600 --> 01:00:34,000 THEY BIND TO THE PARTNER OR 1222 01:00:34,000 --> 01:00:35,600 SOMETHING LIKE THAT. 1223 01:00:35,600 --> 01:00:36,560 YEAH. 1224 01:00:36,560 --> 01:00:39,000 IN PRINCIPLE YOU COULD IMAGINE 1225 01:00:39,000 --> 01:00:42,440 THAT FOR CHAPERONES -- THE 1226 01:00:42,440 --> 01:00:45,520 REGIONS THAT ARE BURIED INSIDE 1227 01:00:45,520 --> 01:00:48,080 THE PROTEIN -- THE PROTEIN 1228 01:00:48,080 --> 01:00:53,240 DOESN'T INTERACT ANY MORE BUT 1229 01:00:53,240 --> 01:00:56,440 WITH TRIC INTERACTS ON THE 1230 01:00:56,440 --> 01:00:57,120 SURFACE. 1231 01:00:57,120 --> 01:01:00,440 HOW IT IS RELEASED IS AN 1232 01:01:00,440 --> 01:01:10,800 INTERESTING QUESTION. 1233 01:01:14,680 --> 01:01:17,160 >> I'M CURIOUS ABOUT SPATIAL 1234 01:01:17,160 --> 01:01:18,480 REGULATIONS AT THE QUALITY 1235 01:01:18,480 --> 01:01:18,840 CONTROL. 1236 01:01:18,840 --> 01:01:21,800 AND NORMALLY WE KNOW THAT THE 1237 01:01:21,800 --> 01:01:24,200 CHAPERONES FUNCTION TO PREVENT 1238 01:01:24,200 --> 01:01:28,880 PROTEIN TO PREVENT AGGREGATION. 1239 01:01:28,880 --> 01:01:31,760 CAN YOU ELABORATE MORE ON HOW 1240 01:01:31,760 --> 01:01:33,600 WHAT IS THE UNDERLYING 1241 01:01:33,600 --> 01:01:36,160 MECHANISM. 1242 01:01:36,160 --> 01:01:39,560 >> SO I DON'T KNOW. 1243 01:01:39,560 --> 01:01:42,280 BUT ONE THING THAT IS VERY COOL 1244 01:01:42,280 --> 01:01:46,120 ABOUT -- PROTEINS IS THEY HAVE 1245 01:01:46,120 --> 01:01:49,640 -- CRYSTALLINE DOMAIN AND THEY 1246 01:01:49,640 --> 01:01:55,160 HAVE LONG -- ---AND WORD FROM 1247 01:01:55,160 --> 01:01:59,120 RACHAEL HAS SHOWN THAT THESE -- 1248 01:01:59,120 --> 01:02:01,560 -- INTERACT WITH THE CRYSTALLINE 1249 01:02:01,560 --> 01:02:03,720 DOMAIN SO THEY FOLD BACK IN AND 1250 01:02:03,720 --> 01:02:07,280 MAYBE ACTIVATE SUBSTRATE BINDING 1251 01:02:07,280 --> 01:02:10,560 SO ONE POSSIBILITIES THAT WHEN 1252 01:02:10,560 --> 01:02:13,080 THE SUBSTRATE BINDS -- THESE 1253 01:02:13,080 --> 01:02:14,720 OPEN UP AND THEY FORM SOME SORT 1254 01:02:14,720 --> 01:02:17,800 OF MeSH WORK SO IT'S NOT 1255 01:02:17,800 --> 01:02:19,080 APPEAR AN AGGREGATE. 1256 01:02:19,080 --> 01:02:21,640 IT'S COMPLETELY SOLUBLE BUT THEY 1257 01:02:21,640 --> 01:02:23,680 FORM A MeSH WORK WITH THE 1258 01:02:23,680 --> 01:02:25,680 SUBSTRATE THAT FORMS SOME SORT 1259 01:02:25,680 --> 01:02:26,880 OF CONDENSATE. 1260 01:02:26,880 --> 01:02:29,160 I HATE THAT WORD BUT SOMETHING 1261 01:02:29,160 --> 01:02:31,920 LIKE THAT. 1262 01:02:31,920 --> 01:02:34,960 -- EXPRESSED ALL OF THE HUMAN 1263 01:02:34,960 --> 01:02:38,640 SMALL -- PROTEINS IN THE 1264 01:02:38,640 --> 01:02:41,040 DELETION AND SHOWED THAT SOME OF 1265 01:02:41,040 --> 01:02:43,040 THE THEM CAN DESTROY THE 1266 01:02:43,040 --> 01:02:44,000 PHENOTYPE. 1267 01:02:44,000 --> 01:02:47,000 THIS COULD BE A GENERAL 1268 01:02:47,000 --> 01:02:51,200 PROPERTY. 1269 01:02:51,200 --> 01:02:52,280 >> WELL THANK YOU VERY MUCH. 1270 01:02:52,280 --> 01:02:54,320 WE HAVE TO CUT IT OFF HERE. 1271 01:02:54,320 --> 01:02:57,640 >> YOU CAN E-MAIL ME YOUR 1272 01:02:57,640 --> 01:02:58,080 QUESTIONS. 1273 01:02:58,080 --> 01:03:00,680 I WILL BE HAPPY TO ANSWER THEM. 1274 01:03:00,680 --> 01:03:01,760 >> THANK YOU. 1275 01:03:01,760 --> 01:03:03,320 IT'S A WONDERFUL TALK. 1276 01:03:03,320 --> 01:03:04,000 THANK YOU VERY MUCH. 1277 01:03:04,000 --> 01:03:14,760 [ APPLAUSE ]