1 00:00:05,117 --> 00:00:06,651 SO WELCOME, EVERYBODY AND THANK 2 00:00:06,651 --> 00:00:08,120 YOU FOR COMING. 3 00:00:08,120 --> 00:00:09,187 AS WE BEGIN PLEASE NOTE THAT 4 00:00:09,187 --> 00:00:12,524 THIS LECTURE IS BEING RECORDED. 5 00:00:12,524 --> 00:00:20,132 I AM JOHN LORSCH AND I WANT TO 6 00:00:20,132 --> 00:00:24,469 WELCOME YOU TO THE 7 00:00:24,469 --> 00:00:27,005 DEWITT LECTURE, HE WAS THE THIRD 8 00:00:27,005 --> 00:00:30,275 DIRECTOR TO NIGMS AND HAD A 9 00:00:30,275 --> 00:00:33,578 STRONG BASIS IN CELLULAR, AND 10 00:00:33,578 --> 00:00:34,413 MOLECULAR CHEMISTRY. 11 00:00:34,413 --> 00:00:36,048 HE SPENT MOST OF HIS CAREER 12 00:00:36,048 --> 00:00:37,449 SERVING AT 2 INSTITUTES AND AS 13 00:00:37,449 --> 00:00:39,217 THE OFFICE OF DIRECTOR OF NIH 14 00:00:39,217 --> 00:00:41,086 AND PARTICULARLY WELL KNOWN AS A 15 00:00:41,086 --> 00:00:42,954 CO-AUTHOR OF THE EARLY ADDITIONS 16 00:00:42,954 --> 00:00:45,991 OF THE TEXTBOOK PRINCIPLES OF 17 00:00:45,991 --> 00:00:46,324 BIOCHEMISTRY. 18 00:00:46,324 --> 00:00:48,060 IN RECOGNITION OF HIS MANY 19 00:00:48,060 --> 00:00:48,693 ACHIEVEMENTS AND CONTRIBUTIONS, 20 00:00:48,693 --> 00:00:50,762 BOTH WITHIN AND OUTSIDE OF NIH, 21 00:00:50,762 --> 00:00:53,131 HE HAS THE RARE DISTINCTION OF 22 00:00:53,131 --> 00:00:58,703 HAVING 2 NIH ENTITIES NAMED IN 23 00:00:58,703 --> 00:01:01,940 HIS HONOR, THIS LECTURE AS WELL 24 00:01:01,940 --> 00:01:11,983 AS THE DEWITT ST ADTMAN 25 00:01:11,983 --> 00:01:12,250 BUILDING. 26 00:01:12,250 --> 00:01:13,752 THE WAY THE LECTURE WORKS IS 27 00:01:13,752 --> 00:01:16,154 THAT THE SPEAKER DR. RY YOUNG 28 00:01:16,154 --> 00:01:17,789 WILL GIVE A 50 MINUTE TALK ON 29 00:01:17,789 --> 00:01:19,324 HIS RESEARCH WHICH WILL BE 30 00:01:19,324 --> 00:01:22,427 FOLLOWED BY A 10 MINUTE QUESTION 31 00:01:22,427 --> 00:01:24,129 AND ANSWER SESSION. 32 00:01:24,129 --> 00:01:27,566 YOU CAN ASK YOUR QUESTIONS BY 33 00:01:27,566 --> 00:01:28,700 USING THE MICROPHONE IN THE ROOM 34 00:01:28,700 --> 00:01:30,535 IF YOU'RE HERE WITH US IN PERSON 35 00:01:30,535 --> 00:01:32,270 OR BY SELECTING THE ACCEPTED 36 00:01:32,270 --> 00:01:35,540 LIVE FEEDBACK FROM THE VIDEOCAST 37 00:01:35,540 --> 00:01:37,109 SCREEN IF YOU ARE YOINING US 38 00:01:37,109 --> 00:01:37,375 VIRTUALLY. 39 00:01:37,375 --> 00:01:39,744 IF YOU ARE SEEKING CONTINUE 40 00:01:39,744 --> 00:01:40,412 MEDICAL EDUCATION CREDIT THE 41 00:01:40,412 --> 00:01:42,147 CODE IS 5 WHERE ARE 098. 42 00:01:42,147 --> 00:01:42,514 LET ME REPEAT. 43 00:01:42,514 --> 00:01:50,489 FIVE WHERE . 44 00:01:50,489 --> 00:01:50,722 5009ATE. 45 00:01:50,722 --> 00:01:53,024 SO TODAY'S SPEAKER RY YOUNG JOIN 46 00:01:53,024 --> 00:01:57,295 US A VERY DISTINGUISHED LIST OF 47 00:01:57,295 --> 00:02:00,932 STADTMAN LECTURERS, HE IS THE 48 00:02:00,932 --> 00:02:02,801 FOUNDER AND DISTINGUISHED 49 00:02:02,801 --> 00:02:05,670 PROFESSOR EMERITUS AT 50 00:02:05,670 --> 00:02:07,806 BIOCHEMISTRY AND BIOPHYSICS AT 51 00:02:07,806 --> 00:02:11,209 TEXAS A&M UNIVERSITY. 52 00:02:11,209 --> 00:02:16,615 HE STUDIED BACTERIAIA PHAGES AND 53 00:02:16,615 --> 00:02:19,718 HOW THEY LYS THE CELLS. 54 00:02:19,718 --> 00:02:21,253 EVEN WHEN OTHERS TURNED AWAY IN 55 00:02:21,253 --> 00:02:24,890 THE FIELD HE SAW THE BENEFIT TO 56 00:02:24,890 --> 00:02:26,491 USING THEM WITH THEIR BIOCHEMIST 57 00:02:26,491 --> 00:02:29,327 RADIOY TO ASK SPECIFIC AND 58 00:02:29,327 --> 00:02:31,329 IMPORTANT BIOLOGICAL QUESTIONS. 59 00:02:31,329 --> 00:02:35,133 DURING TODAY'S LECTURE, RY WILL 60 00:02:35,133 --> 00:02:36,434 DISCUSS HOW THEY CONTROL LYSIS 61 00:02:36,434 --> 00:02:39,404 IN A TIMELY FASHION AND HOW THEY 62 00:02:39,404 --> 00:02:42,374 MIGHT BE HARVESTED FOR 63 00:02:42,374 --> 00:02:43,808 DEVELOPING NEW ANTIBIOTIC ANDS 64 00:02:43,808 --> 00:02:44,376 THERAPY. 65 00:02:44,376 --> 00:02:47,145 HE RECEIVED HIS Ph.D. IN ON 66 00:02:47,145 --> 00:02:48,079 MOLECULAR BIOLOGY FROM THE 67 00:02:48,079 --> 00:02:50,282 UNIVERSITY OF TEXAS AT DALLAS 68 00:02:50,282 --> 00:02:53,351 ALSO A VETERAN OF THE NAVY AND I 69 00:02:53,351 --> 00:02:57,389 THINK HE PROBABLY IS THE 70 00:02:57,389 --> 00:02:58,456 STADTMAN LECTURE AND WALS 71 00:02:58,456 --> 00:03:00,792 LECTURE WHO HAS THE MOST 72 00:03:00,792 --> 00:03:02,160 IMPRESSIVE JOURNEY FOR HOW HE 73 00:03:02,160 --> 00:03:03,361 GOT HERE, HE DIDN'T FLY, HE 74 00:03:03,361 --> 00:03:04,329 DIDN'T TAKE A TRAIN. 75 00:03:04,329 --> 00:03:07,032 HE ACTUALLY WITH HIS WIFE, DROVE 76 00:03:07,032 --> 00:03:10,402 OR PILOTED HIS BOAT ALL THE WAY 77 00:03:10,402 --> 00:03:14,139 FROM TEXAS, THROUGH THE GULF, 78 00:03:14,139 --> 00:03:16,107 THROUGH FLORIDA, UP THE WATER 79 00:03:16,107 --> 00:03:20,645 WAYS INTO THE CHESAPEAKE BAY. 80 00:03:20,645 --> 00:03:21,713 [ APPLAUSE ] 81 00:03:21,713 --> 00:03:22,681 SO THAT IS SOME SERIOUS 82 00:03:22,681 --> 00:03:24,282 DEDICATION TO GET HERE. 83 00:03:24,282 --> 00:03:25,584 SO THANKS FOR COMING RY. 84 00:03:25,584 --> 00:03:29,487 IT'S IMRAIT --GREAT TO HAVE YOU 85 00:03:29,487 --> 00:03:30,555 WITH US. 86 00:03:30,555 --> 00:03:32,224 >> SO FIRST OF ALL, I'M SORRY 87 00:03:32,224 --> 00:03:33,625 ABOUT WEARING MY GLASSES LIKE 88 00:03:33,625 --> 00:03:35,694 THIS, I'M NOT TRYING TO IMITATE 89 00:03:35,694 --> 00:03:38,029 THE OLD FUDY DUDY PROFESSOR BUT 90 00:03:38,029 --> 00:03:41,933 I LOST MY BI FOCALS, MY WIFE 91 00:03:41,933 --> 00:03:43,468 SAYS I RIDICULOUS, I HOPE THEY 92 00:03:43,468 --> 00:03:44,236 DON'T FALL OFF. 93 00:03:44,236 --> 00:03:46,538 THIS SLIDE IS TO MAKE SURE I 94 00:03:46,538 --> 00:03:47,839 ACKNOWLEDGE PEOPLE WHO HAVE DONE 95 00:03:47,839 --> 00:03:48,773 THE WORK, SEVERAL HERE IN THE 96 00:03:48,773 --> 00:03:51,977 BUILDING AND I WILL COME BACK TO 97 00:03:51,977 --> 00:03:52,944 THE VERY END BECAUSE THIS WAS 98 00:03:52,944 --> 00:03:56,248 KIND OF THE WORK PRODUCT, THIS 99 00:03:56,248 --> 00:03:57,616 NIGMS GRANT FUNDED THIS WORK FOR 100 00:03:57,616 --> 00:03:59,284 OVER 40 YEARS AND PART OF 101 00:03:59,284 --> 00:04:00,285 PRODUCT IS ACCIDENT WELL TRAINED 102 00:04:00,285 --> 00:04:03,855 PEOPLE HAVE TBOAN ON TO 103 00:04:03,855 --> 00:04:05,624 SUCCESSIVE SCIENCE IN MANY WAYS. 104 00:04:05,624 --> 00:04:10,161 THE 1S IN RED ARE MEMBERS OF THE 105 00:04:10,161 --> 00:04:11,363 BIOTECH COMPANY AND ALMOST ALL 106 00:04:11,363 --> 00:04:14,466 THE OTHER 1S ARE ON THEIR WAY, 107 00:04:14,466 --> 00:04:16,334 PHAGE BIOLOGY IS A GREAT SYSTEM 108 00:04:16,334 --> 00:04:17,836 FOR TRAINING BECAUSE CAN YOU GET 109 00:04:17,836 --> 00:04:19,537 SPRMS DONE QUICKLY, NOBODY THERE 110 00:04:19,537 --> 00:04:21,873 IS STILL WORKING ON PHAGE LYSIS 111 00:04:21,873 --> 00:04:23,174 OR EACH PHAGE OF THE IT'S BEEN A 112 00:04:23,174 --> 00:04:25,844 GOOD SYSTEM IN THAT REGARD. 113 00:04:25,844 --> 00:04:28,713 OKAY, SO IT'S A GREAT HONOR TO 114 00:04:28,713 --> 00:04:31,016 BE OVER 40 YEARS AGO BEFORE THIS 115 00:04:31,016 --> 00:04:31,983 STARTED THERE'S TILL NOT BEEN AS 116 00:04:31,983 --> 00:04:34,052 FAR AS I CAN TELL, I'M THE FIRST 117 00:04:34,052 --> 00:04:36,621 PHAGE GUY, WHICH IS REALLY WEIRD 118 00:04:36,621 --> 00:04:42,794 SINCE NIGMS IS THE FIRST PLACE 119 00:04:42,794 --> 00:04:44,863 TO GIVE A FAINL TALK, THE FITTER 120 00:04:44,863 --> 00:04:46,131 1 WAS MARK [INDISCERNIBLE] BUT 121 00:04:46,131 --> 00:04:48,533 HE MIGHT HAVE BEEN TALKING ABOUT 122 00:04:48,533 --> 00:04:50,068 YEAST TRANSCRIPTION SO WE'RE 123 00:04:50,068 --> 00:04:51,136 WRITING A HISTORICAL WRONG HERE. 124 00:04:51,136 --> 00:04:53,571 ALL RIGHT, SO I CAN REMEMBER 125 00:04:53,571 --> 00:04:55,106 THIS, IS A RECONSTRUCTION, NOT 126 00:04:55,106 --> 00:04:59,844 THE ACTUAL, BUT BACK IN 1969 OR 127 00:04:59,844 --> 00:05:01,513 70, NO IT WAS EARLIER THAN THAT, 128 00:05:01,513 --> 00:05:03,615 IT WAS LATER THAN THAT IT WAS 129 00:05:03,615 --> 00:05:06,818 72, I THINK, I WAS STANDING IN 130 00:05:06,818 --> 00:05:08,753 JOHN [INDISCERNIBLE]'S LAB AT 131 00:05:08,753 --> 00:05:10,388 HARVARD MEDICAL SCHOOL,IME 5 132 00:05:10,388 --> 00:05:13,892 YEARS OLD, OLD FUDY DUDYS HAVE 133 00:05:13,892 --> 00:05:14,626 TROUBLE WITH DECADES. 134 00:05:14,626 --> 00:05:19,531 ANY RATE, I BORROWED HIS FLASK 135 00:05:19,531 --> 00:05:22,667 AND HIS SPEC TOM TERTO FOLLOW 136 00:05:22,667 --> 00:05:23,601 LAMDA INDUCTIONS, I WON'T TELL 137 00:05:23,601 --> 00:05:26,805 YOU WHY I WAS DOING THESE LAMDA 138 00:05:26,805 --> 00:05:28,206 INDUCTIONS BUT YOU CAN HEAT 139 00:05:28,206 --> 00:05:29,607 SHOCK THE BACTERIA IN A CULT IRB 140 00:05:29,607 --> 00:05:33,378 FLASK AND THEN THE PROPHAGE WILL 141 00:05:33,378 --> 00:05:35,580 SUDDENLY INDUCE AND YOU GET 142 00:05:35,580 --> 00:05:36,514 SYNCHRONOUS INFECTION THIS IS 143 00:05:36,514 --> 00:05:36,815 WAY. 144 00:05:36,815 --> 00:05:38,283 AND THAT'S THE HIGH VALUE. 145 00:05:38,283 --> 00:05:39,617 SO I STARTED 5 CULTURES, I 146 00:05:39,617 --> 00:05:41,386 LOOKED DOWN AND DISCOVERED, 3 147 00:05:41,386 --> 00:05:42,887 MINUTES AGO THAT I HAD NOT 148 00:05:42,887 --> 00:05:48,093 STARTED 1 OF THE CULTURES SO I 149 00:05:48,093 --> 00:05:50,729 SHIFTED IT, SO 1 OF THESE TARTED 150 00:05:50,729 --> 00:05:52,263 LATER, SO IF YOU LOOK AT IT ALL 151 00:05:52,263 --> 00:05:57,135 THE CELLS IN THE ORIGINAL FLASK 152 00:05:57,135 --> 00:06:00,672 HAVE LYSED AND NONE OF THE IN 153 00:06:00,672 --> 00:06:05,009 THE ORIGINAL HAVE LYRK SED AND 154 00:06:05,009 --> 00:06:07,779 IT IS ENACTED BY LAMDA, IT'S 155 00:06:07,779 --> 00:06:08,847 HIGHLY CONTROLLED IN TERMS OF 156 00:06:08,847 --> 00:06:11,282 TIME, SO THIS IS A--THIS IS AN 157 00:06:11,282 --> 00:06:12,684 EVENT THAT CAN KEEP TIME ON THE 158 00:06:12,684 --> 00:06:15,453 ORDER OF LOW MINUTES TO SECONDS. 159 00:06:15,453 --> 00:06:16,955 AND I'VE HAD CHILDREN KEEP TRACK 160 00:06:16,955 --> 00:06:19,591 OF TIME AND DAYS. 161 00:06:19,591 --> 00:06:25,697 THIS TURNS OUT ALL TO BE DUE 162 00:06:25,697 --> 00:06:27,332 SINGLE PROTEIN, THIS LECTURE, BY 163 00:06:27,332 --> 00:06:29,334 THE WAY HAD IS A LECTURE SO YOU 164 00:06:29,334 --> 00:06:33,638 WILL HEAR MOSTLY MY THEORIES ON 165 00:06:33,638 --> 00:06:35,173 WHAT IS HAPPENING OVER SOMETHING 166 00:06:35,173 --> 00:06:36,441 THAT HAS NO CONTROLS AND THEN I 167 00:06:36,441 --> 00:06:38,543 WILL TRY TO COVER THE 168 00:06:38,543 --> 00:06:39,377 ENTIRE--HOW MY UNDERSTANDING IS 169 00:06:39,377 --> 00:06:40,678 OF THE ENTIRE SYSTEM THAT MAKES 170 00:06:40,678 --> 00:06:51,122 GOING FROM THERE TO THERE. 171 00:06:53,825 --> 00:06:55,560 LEGALITYEE SEE IF I CAN NOT 172 00:06:55,560 --> 00:06:55,994 CRASH THE COMPUTER. 173 00:06:55,994 --> 00:06:57,462 THIS IS WHAT THAT LOOKS LIKE AT 174 00:06:57,462 --> 00:06:58,530 THE SINGLE CELL LEVEL AND THIS 175 00:06:58,530 --> 00:06:59,831 IS ALL IN SECONDS HERE SO WE 176 00:06:59,831 --> 00:07:02,167 LOOK AT IT IN MILLISECOND FUEL 177 00:07:02,167 --> 00:07:03,234 AND NOTICE WHAT'S GOING ON, 178 00:07:03,234 --> 00:07:04,536 NOTICE IT'S NOT JUST A GENERAL 179 00:07:04,536 --> 00:07:06,738 BLOW OUT OF THE WHOLE THING, 180 00:07:06,738 --> 00:07:08,039 THERE'S A SPECIFIC LESION THAT 181 00:07:08,039 --> 00:07:09,007 OCCURS ISSUES AT 1 POINT IN THE 182 00:07:09,007 --> 00:07:10,608 CELL, WE CALL THIS LOCAL BLOW 183 00:07:10,608 --> 00:07:12,210 OUT SO WHATEVER THAT TIMING 184 00:07:12,210 --> 00:07:14,879 EVENT THAT WE LOOK AT THE 185 00:07:14,879 --> 00:07:16,047 MACROSCOPIC LEVEL OCCURS ALSO AT 186 00:07:16,047 --> 00:07:18,149 THE SINGLE CELL LEVEL, THIS 187 00:07:18,149 --> 00:07:20,084 HAPPENS AT A SINGLE PLACE AT A 188 00:07:20,084 --> 00:07:21,286 SINGLE TIME AND FOR SEVERAL 189 00:07:21,286 --> 00:07:21,920 SECURITIZATION.S AFTER THAT THE 190 00:07:21,920 --> 00:07:23,188 REST OF THE STRUCTURE OF THE 191 00:07:23,188 --> 00:07:25,857 CELL IS NORMAL AND THAT FLOW 192 00:07:25,857 --> 00:07:26,491 OUT, PRESUMABLY ALTHOUGH WE 193 00:07:26,491 --> 00:07:28,793 CAN'T SEE IT, ALL THE HUNDRED OR 194 00:07:28,793 --> 00:07:29,894 SO PHAGE PARPARTICLES ARE 195 00:07:29,894 --> 00:07:30,829 EXPELLED FROM THAT END OF THE 196 00:07:30,829 --> 00:07:32,664 CELL IN THIS CASE, ABOUT 80% OF 197 00:07:32,664 --> 00:07:37,435 THE TIME IT DOES OCCUR AT THE 198 00:07:37,435 --> 00:07:38,336 POLL. 199 00:07:38,336 --> 00:07:40,905 THIS DOES DONE BY 200 00:07:40,905 --> 00:07:41,439 JCERTAINLY--CERTAINLY SSE 201 00:07:41,439 --> 00:07:46,144 CAHILL, WHO HAS HIS OWN LAB AT 202 00:07:46,144 --> 00:07:46,377 SANDIA. 203 00:07:46,377 --> 00:07:47,212 SOIME PRETTY SURE THAT 204 00:07:47,212 --> 00:07:47,946 [INDISCERNIBLE] WAS THERE WHEN 205 00:07:47,946 --> 00:07:49,914 THEY DID THIS, THIS IS NOT 206 00:07:49,914 --> 00:07:52,550 LAMDA, THIS IS T4 SO IT'S 207 00:07:52,550 --> 00:07:56,454 UNUSUAL FOR ME TO SHOW T4 SLIDES 208 00:07:56,454 --> 00:08:04,462 I'M A LAMDA TRIBE PERSON BUT THE 209 00:08:04,462 --> 00:08:05,196 DELBRUCK GROUP DID THIS 210 00:08:05,196 --> 00:08:08,366 EXPERIMENT, AND NOTICE THAT VERY 211 00:08:08,366 --> 00:08:09,901 SHARP TEMPERRAL CONTROL OF LYSIS 212 00:08:09,901 --> 00:08:10,935 EXISTS ISSUES IT'S IN A 213 00:08:10,935 --> 00:08:12,537 DIFFERENT TIME BECAUSE IT'S A 214 00:08:12,537 --> 00:08:15,240 DIFFERENT PHAGE BUT IT'S VERY 215 00:08:15,240 --> 00:08:17,909 ABRUPT AND SO IT'S THE GRAPHICAL 216 00:08:17,909 --> 00:08:20,545 REPRESENTATION OF WHAT WE 217 00:08:20,545 --> 00:08:21,246 ALREADY SAW. 218 00:08:21,246 --> 00:08:23,314 SO IF YOU-1 OTHER THING WE HAVE 219 00:08:23,314 --> 00:08:24,482 TO EXPLAIN IS THE UNIVERSAL 220 00:08:24,482 --> 00:08:26,317 CHARACTER OF PHAGE INDUCED LYSIS 221 00:08:26,317 --> 00:08:28,653 AND THAT IS IF YOUADAD CYANIDE 222 00:08:28,653 --> 00:08:30,388 EARLY IN INFECTION, IT JUST 223 00:08:30,388 --> 00:08:31,723 STOPS ALL ENEMBRYONIC STEMY 224 00:08:31,723 --> 00:08:32,557 METABOLISM AND NOTHING HAPPENS 225 00:08:32,557 --> 00:08:35,426 IF YOU LOOK INDEED THIS CELL, 226 00:08:35,426 --> 00:08:36,895 PHAGES AREN'T REPLICATING, NO 227 00:08:36,895 --> 00:08:38,196 ASSEMBLY IS GOING ON, NOTHING'S 228 00:08:38,196 --> 00:08:39,597 HAPPENING AND THAT SHOWS UP AS 229 00:08:39,597 --> 00:08:41,132 NO CHANGE IN O. D. FOR A LONG 230 00:08:41,132 --> 00:08:43,001 TOWARD OF TIME BUT IF YOU WAIT, 231 00:08:43,001 --> 00:08:45,737 SAY 15 OR 20 MINUTES AND NOW, 232 00:08:45,737 --> 00:08:48,339 INSIDE THIS CELL PHAGE PARTICLES 233 00:08:48,339 --> 00:08:51,376 ARE BEING ASSEMBLED, ROBUST GENE 234 00:08:51,376 --> 00:08:52,877 EXPRESSION, THE LYSISSED 235 00:08:52,877 --> 00:08:54,579 PROTEINS ARE BEING EXPRESSED, 236 00:08:54,579 --> 00:08:56,347 THE INSTANT YOU ADD ANYTHING 237 00:08:56,347 --> 00:08:58,216 ELSE THAT CAN DEPOLARIZE THE 238 00:08:58,216 --> 00:08:59,183 MEMBRANE, YOU TRIGGER LYSIS 239 00:08:59,183 --> 00:09:01,085 IMMEDIATELY AND THIS IS CALLED 240 00:09:01,085 --> 00:09:04,422 PREMATURE LYSIS, THIS WAS THAT 241 00:09:04,422 --> 00:09:12,330 TERMINOLOGY INVENTED BY MAX 242 00:09:12,330 --> 00:09:14,098 DELBRUCK, SO IT'S A PHENOMENON, 243 00:09:14,098 --> 00:09:15,466 WHEN WE TRY TO FIGURE OUT HOW 244 00:09:15,466 --> 00:09:20,104 THEY DO IT AND WE HAVE TO 245 00:09:20,104 --> 00:09:21,406 EXPLAIN TO EXPLAIN THE TIMING 246 00:09:21,406 --> 00:09:23,274 AND IN ADDITION YOU CAN DISRUPT 247 00:09:23,274 --> 00:09:25,610 THAT BY COLLAPSING THE MEMBRANE 248 00:09:25,610 --> 00:09:27,211 POTENTIAL, I'M SORRY, I'M SO 249 00:09:27,211 --> 00:09:32,550 HORSE, I'VE ANSWERED SO MANY 250 00:09:32,550 --> 00:09:32,917 QUESTIONS TODAY. 251 00:09:32,917 --> 00:09:36,621 OKAY, SO, THE MODEL HAS TO 252 00:09:36,621 --> 00:09:37,288 EXPLAIN BOTH THINGS. 253 00:09:37,288 --> 00:09:41,960 ALL RIGHT, SO THIS IS THE LAMDA 254 00:09:41,960 --> 00:09:42,226 LYSIS SET. 255 00:09:42,226 --> 00:09:44,629 SO IF YOU DRAW TO SCALE THIS IS 256 00:09:44,629 --> 00:09:47,699 AT THE RIGHT END OF LAMDA. 257 00:09:47,699 --> 00:09:49,534 IS THIS WORKING IN YEAH. 258 00:09:49,534 --> 00:09:51,302 SO THERE'S THE LATE PROMOTER AND 259 00:09:51,302 --> 00:09:53,137 THE FIRST GENES ARE THE LYSIS 260 00:09:53,137 --> 00:09:54,439 YEENS THAT ARE COUNTER ENTUMOR 261 00:09:54,439 --> 00:09:55,707 SPECTRUMMATIVE AND THEN THERE'S 262 00:09:55,707 --> 00:09:57,008 THE HEAD YEENS AND TAIL GENES 263 00:09:57,008 --> 00:09:58,242 AND THIS WOULD BE ALL THE WAY IN 264 00:09:58,242 --> 00:10:01,613 THE NEXT BUILDING FOR THE NEXT 265 00:10:01,613 --> 00:10:03,948 20 YEARS BUT FROM 1 TRANSCRIPT, 266 00:10:03,948 --> 00:10:06,184 IT TURNS OUT AFTER 8 MINUTES OF 267 00:10:06,184 --> 00:10:08,286 INFECTION, AND IT MAKE ITS EASY 268 00:10:08,286 --> 00:10:10,054 IN TERMS OF KINETICS BECAUSE 269 00:10:10,054 --> 00:10:11,556 THERE'S NOT A COMPLEX 270 00:10:11,556 --> 00:10:12,957 TRANSCRIPTIONAL O THIS IS EITHER 271 00:10:12,957 --> 00:10:14,692 ON OR OFF AND AFTER THAT IT'S A 272 00:10:14,692 --> 00:10:17,362 MATTER OF COPY NUMBER AND THERE 273 00:10:17,362 --> 00:10:21,032 ARE 4 GENES, SR, RZ, AND RZ1. 274 00:10:21,032 --> 00:10:22,233 WE WILL SPEND MOST OF OUR TIME 275 00:10:22,233 --> 00:10:23,968 TALKING ABOUT THE GENES ARRANGED 276 00:10:23,968 --> 00:10:29,073 BY THE WAY IN THE ORDER OF 277 00:10:29,073 --> 00:10:31,576 ASSAULTING THE ENVELOPE. 278 00:10:31,576 --> 00:10:36,280 SO THE FIRST GENE, GENE S, GENE 279 00:10:36,280 --> 00:10:38,516 S ENCODES 2 PROTEEPS. 280 00:10:38,516 --> 00:10:40,585 IT'S 107 CODONS LONG, MAKES 1 281 00:10:40,585 --> 00:10:43,988 PROTEIN FULL LENGTH AND THAT'S 282 00:10:43,988 --> 00:10:47,258 THE ACTUAL ANTIHOLEN AND MAKES A 283 00:10:47,258 --> 00:10:47,892 SHORTER BY EARLIERINNISHIATION 284 00:10:47,892 --> 00:10:48,993 AND THAT WILL BE THE HOLE AND 285 00:10:48,993 --> 00:10:50,962 THAT WILL BE THE START OF THE 286 00:10:50,962 --> 00:10:52,730 SHOW, WE WON'T TALK ABOUT HOW 287 00:10:52,730 --> 00:10:55,133 THIS WORKS BUT YOU CAN TELL FROM 288 00:10:55,133 --> 00:10:58,269 ITS NAME THAT IT WORKS IN 289 00:10:58,269 --> 00:10:59,103 DIRECTLY INHIBITING THE HOLE. 290 00:10:59,103 --> 00:11:00,505 WE WILL TALK ABOUT THIS AT THE 291 00:11:00,505 --> 00:11:03,141 END OF THE SEMINAR, IT THE NEXT 292 00:11:03,141 --> 00:11:05,043 GENE IS THE ENDOLIESIN AND THE 293 00:11:05,043 --> 00:11:07,011 LYSOSIEM, IT'S THE ENZYME THAT 294 00:11:07,011 --> 00:11:11,215 DEGRADES THE CELL WALL AND OF 295 00:11:11,215 --> 00:11:12,950 COURSE IT ATTACKS THE PEPTIDE O 296 00:11:12,950 --> 00:11:15,286 O GLYCAN AND THEN THE 2 GENES IN 297 00:11:15,286 --> 00:11:19,791 THE SAME DNA, RZ, AND RZ1, 153 298 00:11:19,791 --> 00:11:22,493 AND 160 CODONS LONG, COMPLETELY 299 00:11:22,493 --> 00:11:23,728 DIFFERENT PROTEINS, BOTH 300 00:11:23,728 --> 00:11:26,230 REQUIRED WHICH RAISES ALL KINDS 301 00:11:26,230 --> 00:11:27,365 OF EVOLUTIONARY QUANDARYS. 302 00:11:27,365 --> 00:11:30,001 THEY'RE ENCODING THE 2 SUBUNITS 303 00:11:30,001 --> 00:11:31,836 OF THE SPANIN PROTEIN AND 304 00:11:31,836 --> 00:11:33,004 THEY'RE REQUIRED FOR DISRUPTING 305 00:11:33,004 --> 00:11:33,638 THE OUTER EMPLOYMENT PROGRAMS 306 00:11:33,638 --> 00:11:38,476 BRAIN AND IT'S ESSENTIAL FOR 307 00:11:38,476 --> 00:11:38,743 LYSIS. 308 00:11:38,743 --> 00:11:40,678 SO IMPORTANT TO NOTE THAT YOU 309 00:11:40,678 --> 00:11:43,114 CAN EITHER USE THE PHAGE OR YOU 310 00:11:43,114 --> 00:11:46,417 CAN CLONE THOSE 4 YEENS INTO AN 311 00:11:46,417 --> 00:11:48,219 INDUCIBLE BACK DROP TO GET THE 312 00:11:48,219 --> 00:11:48,653 SAME PHENOTYPES. 313 00:11:48,653 --> 00:11:50,154 YOU CHANGE THE TYPES JUST BASED 314 00:11:50,154 --> 00:11:51,289 ON COPY NUMBER BUT EVERYTHING 315 00:11:51,289 --> 00:11:53,858 ELSE IT DOESN'T MATTER IF 316 00:11:53,858 --> 00:11:54,792 YOU'RE--THERE'S NO OTHER YEENS 317 00:11:54,792 --> 00:11:56,394 IN THE PHAGE THAT HAVE ANY 318 00:11:56,394 --> 00:11:57,595 EFFECT ON FUNCTION OR TIMING. 319 00:11:57,595 --> 00:11:59,330 SO SOMETIMES I WILL SHOW YOU 320 00:11:59,330 --> 00:12:00,631 DATA FROM PLASMA AND IT WON'T 321 00:12:00,631 --> 00:12:04,302 EVEN TELL YOU WHEN I'M DOING 322 00:12:04,302 --> 00:12:04,502 THAT. 323 00:12:04,502 --> 00:12:06,270 OKAY, THE FIRST THING, TO DO IS 324 00:12:06,270 --> 00:12:10,007 TO GET RID OF THE LAMDA 325 00:12:10,007 --> 00:12:12,410 LYSOSIEM, THIS IS THE BASIC 326 00:12:12,410 --> 00:12:17,081 STRUCTURE OF THE BACTERIAL CELL 327 00:12:17,081 --> 00:12:19,417 WALL, AND ALL YOU NEED TO KNOW 328 00:12:19,417 --> 00:12:21,452 IS AMONG THE 4 MOST IMPORTANT 329 00:12:21,452 --> 00:12:23,654 SCIENTIFIC TRIBES, THE T7 T4 330 00:12:23,654 --> 00:12:26,157 LAMDA AND T5 TRIBES, EACH 1 OF 331 00:12:26,157 --> 00:12:29,360 THEM HAD A DIFFERENT GENE CALLED 332 00:12:29,360 --> 00:12:30,695 LYSOSIEM, AND SO IT TURNS OUT 333 00:12:30,695 --> 00:12:33,631 THAT ALL 4 OF THESE LYSOSIEMS 334 00:12:33,631 --> 00:12:34,699 ARE COMPLETELY DIFFERENT 335 00:12:34,699 --> 00:12:39,570 ENZYMES, SO LAMDA MAKES A 336 00:12:39,570 --> 00:12:42,273 TRANSGLYCOSYLASE AND T4 MAKES A 337 00:12:42,273 --> 00:12:44,575 GLUE MARIOUS COSDACE O IT'S A 338 00:12:44,575 --> 00:12:46,644 BOND, IT MAKES THE BOND BETWEEN 339 00:12:46,644 --> 00:12:50,281 THE PEPTIDE IS THE CELL WALL AND 340 00:12:50,281 --> 00:12:54,085 THE TBLIEKOCIDE, AND THEN T5 341 00:12:54,085 --> 00:12:54,852 MAKES AN ENDOPEPTIDE O 342 00:12:54,852 --> 00:12:58,055 EDUCATIONAL ACE, AND THEY'RE 343 00:12:58,055 --> 00:13:00,358 THEY'RE NOT MUCH INTEREST, 344 00:13:00,358 --> 00:13:01,759 THEY'RE INTERCHANGEABLE, THEY'RE 345 00:13:01,759 --> 00:13:04,695 ALWAYS PRODUCED IN 100 FOLD 346 00:13:04,695 --> 00:13:05,163 ACCESS. 347 00:13:05,163 --> 00:13:06,664 AND THEY'RE INTERCHANGEABLE AND 348 00:13:06,664 --> 00:13:08,566 THEY'RE BORING, SO THE--THE 349 00:13:08,566 --> 00:13:09,467 INTERESTING GIVING--YOUS HERE 350 00:13:09,467 --> 00:13:11,636 ARE THE WHOLE 1S. 351 00:13:11,636 --> 00:13:17,408 SO I MADE THE WORD UP, HOLEN, IF 352 00:13:17,408 --> 00:13:18,910 EVERYBODY GENERATES A WORD 353 00:13:18,910 --> 00:13:20,845 THAT'S TOO MUCH ADDITIONS TO THE 354 00:13:20,845 --> 00:13:23,347 ENGLISH LANGUAGE. 355 00:13:23,347 --> 00:13:26,083 BUT SO LAMDA IS TO EMPHASIZE THE 356 00:13:26,083 --> 00:13:28,286 FACT THAT THEY ALL HAVE AT LEAST 357 00:13:28,286 --> 00:13:29,353 1 TRANSMEMBRANE DOMAIN WHICH IS 358 00:13:29,353 --> 00:13:30,421 NOT SURPRISING IF IT'S DOING 359 00:13:30,421 --> 00:13:32,623 SOMETHING FOR THE INNER 360 00:13:32,623 --> 00:13:35,393 MEMBRANE, AND THEN, SO LAMDA, 361 00:13:35,393 --> 00:13:37,562 HAS 3 TRANSMEMBRANE DOMAINS, 362 00:13:37,562 --> 00:13:39,463 IT'S AMINO TERMINUS IS OUT AND 363 00:13:39,463 --> 00:13:42,200 THE C-TERMINUS IS IN, AND HAD IS 364 00:13:42,200 --> 00:13:44,202 THE TOPOLOGY, THE OTHER--THE 365 00:13:44,202 --> 00:13:46,404 OTHER 4 TOPOLOGYS, THESE ARE THE 366 00:13:46,404 --> 00:13:48,706 ONLY 4 TOPOLOGYYS THAT HAVE BEEN 367 00:13:48,706 --> 00:13:49,774 EXPERIMENTALLY CONFIRMED SO 368 00:13:49,774 --> 00:13:52,210 THERE'S TYPE 1, 2, 3, 4, AND IF 369 00:13:52,210 --> 00:13:53,444 WE HAD BEEN SMART THIS WOULD BE 370 00:13:53,444 --> 00:13:54,912 TYPE 1 CAN THAT WOULD BE TYPE 3 371 00:13:54,912 --> 00:13:55,746 BECAUSE THEN THE TYPE NUMBER 372 00:13:55,746 --> 00:13:57,615 WOULD BE THE NUMBER OF 373 00:13:57,615 --> 00:13:58,549 TRANSMEMBRANE DOMAINS. 374 00:13:58,549 --> 00:14:00,284 BUT WE DIDN'T DO IT SMARTLY SO 375 00:14:00,284 --> 00:14:01,719 TYPE 1 WILL HAVE 3 AND THE LAST 376 00:14:01,719 --> 00:14:04,822 TIME I LOOKED WHICH WAS LAST 377 00:14:04,822 --> 00:14:08,593 WEEK, THERE ARE 215,000 RECORDS 378 00:14:08,593 --> 00:14:09,994 IN THE IDENTICAL PROTEIN GROUP 379 00:14:09,994 --> 00:14:12,630 AND MY EXPERTISE IS NOT 380 00:14:12,630 --> 00:14:14,265 ENVIRONMENTAL PHYSICS BUT I 381 00:14:14,265 --> 00:14:15,600 LOOKEDDA THE FIRST 600 OR SO AND 382 00:14:15,600 --> 00:14:16,100 THEY'RE ALL DIFFERENT. 383 00:14:16,100 --> 00:14:18,069 SO THERE ARE THOUSANDS, AT LEAST 384 00:14:18,069 --> 00:14:19,270 THOUSANDS OF UNRELATED GENE 385 00:14:19,270 --> 00:14:21,439 FAMILIES AND THEY ALL HAVE AT 386 00:14:21,439 --> 00:14:23,074 LEAST 1 TRANSMEMBRANE DOMAIN BUT 387 00:14:23,074 --> 00:14:26,143 NOTICE, 2 THAT 1 OF THEM WHICH 388 00:14:26,143 --> 00:14:29,213 IS THE T-4 WHICH IS ALSO SHARED 389 00:14:29,213 --> 00:14:33,251 BY T-5 HAS A PERIPLAZ MIDSIC 390 00:14:33,251 --> 00:14:34,585 DOMAINS OF ABOUT AMINO ACIDS AND 391 00:14:34,585 --> 00:14:35,887 HAD IS A SIGNATURES 95 392 00:14:35,887 --> 00:14:36,520 TRANSDUCTION RECEPTOR AND THIS 393 00:14:36,520 --> 00:14:39,056 IS THE ONLY CASE WE KNOW IN 394 00:14:39,056 --> 00:14:40,057 BIOLOGY WHERE EXTERNAL 395 00:14:40,057 --> 00:14:42,293 INFORMATION IS PUT INTO LYSIS 396 00:14:42,293 --> 00:14:42,960 TIMING, UNFORTUNATELY, WE'RE NOT 397 00:14:42,960 --> 00:14:44,228 GOING TO BE ABLE TO TALK ABOUT 398 00:14:44,228 --> 00:14:46,931 THIS UNLESS I GET A PREGNANT 399 00:14:46,931 --> 00:14:49,400 QUESTION NEAR THE END BUT IT'S A 400 00:14:49,400 --> 00:14:50,868 WONDERFUL STORY THAT'S RECENTLY 401 00:14:50,868 --> 00:14:52,703 MADE PROGRESS AT THE STRUCTURAL 402 00:14:52,703 --> 00:14:54,038 LEVEL, BUT IT'S NOT RELATED TO 403 00:14:54,038 --> 00:15:02,280 THE BASIC STORY OF HOW THE 404 00:15:02,280 --> 00:15:03,114 TIMING OCCURS. 405 00:15:03,114 --> 00:15:04,782 SO THIS IS ONLY 1 OF THE 2 406 00:15:04,782 --> 00:15:06,951 PROTEINS MADE BY THE S-GENE, 407 00:15:06,951 --> 00:15:07,919 WHICH IS THE TRUE 408 00:15:07,919 --> 00:15:10,187 [INDISCERNIBLE]. 409 00:15:10,187 --> 00:15:12,223 SO THE INITIAL--BACK WHEN I WAS 410 00:15:12,223 --> 00:15:14,558 STARTING TO GET MY LAB TOGETHER 411 00:15:14,558 --> 00:15:17,495 I GOT A COUPLE PAPERS IN GRANT 412 00:15:17,495 --> 00:15:19,697 REVIEW WAS THAT THE SIMPLEST 413 00:15:19,697 --> 00:15:22,400 IDEA WAS KNOWN THAT CYANIDE 414 00:15:22,400 --> 00:15:25,036 COULD CAUSE PHAGE INFECTIONS, 415 00:15:25,036 --> 00:15:28,673 YOU GIVE CYANIDE, EVERYTHING 416 00:15:28,673 --> 00:15:29,340 COLLAPSES, EVERYTHING LYSIS, 417 00:15:29,340 --> 00:15:33,411 THAT WAS ALREADY KNOWN, SO 418 00:15:33,411 --> 00:15:36,047 HOLEIN CONTRIBUTE OUT THE 419 00:15:36,047 --> 00:15:38,482 MEMBRANE, IF YOU EXPRESS 420 00:15:38,482 --> 00:15:42,520 MEMBRANE PROTEINS YOU POISON THE 421 00:15:42,520 --> 00:15:43,487 PROTEIN, AND WHATEVER WAS 422 00:15:43,487 --> 00:15:44,789 HAPPENING WAS HAPPENING THE SAME 423 00:15:44,789 --> 00:15:47,525 AND NORMAL INDUCTION HAS A 424 00:15:47,525 --> 00:15:48,125 PRACTICES MATURE LYSIS. 425 00:15:48,125 --> 00:15:49,060 SO WE ANSWERED THAT QUESTION 426 00:15:49,060 --> 00:15:50,227 QUITE A LONG TIME AGO. 427 00:15:50,227 --> 00:15:53,731 THIS IS WORK DONE BY 428 00:15:53,731 --> 00:15:54,799 [INDISCERNIBLE] FROM THE COLLEGE 429 00:15:54,799 --> 00:15:56,434 OF LONDON AND COLLABORATOR WITH 430 00:15:56,434 --> 00:16:00,137 MIKE [INDISCERNIBLE] AT TEXAS 431 00:16:00,137 --> 00:16:00,304 A&M. 432 00:16:00,304 --> 00:16:01,572 LET'SEE IF I CAN DO THIS, 433 00:16:01,572 --> 00:16:03,207 IMAGINE THESE ARE CELLS THAT ARE 434 00:16:03,207 --> 00:16:06,243 TETHERED TO THE MEMBRANE BY 435 00:16:06,243 --> 00:16:08,546 THEIR FLAGELLA TO TELL IT TO THE 436 00:16:08,546 --> 00:16:09,280 FLAGELLA AND BEGINNING OF THE 437 00:16:09,280 --> 00:16:11,916 DIRECT READ OUT OF THE MEMBRANE 438 00:16:11,916 --> 00:16:13,117 POTENTIAL BECAUSE THE FLAGELLA, 439 00:16:13,117 --> 00:16:14,752 BY THE WAY, THESE ARE LOCK 440 00:16:14,752 --> 00:16:16,087 INDEED A CLOCKWISE ORIENTATION 441 00:16:16,087 --> 00:16:19,323 SO WE KNOW THEIR PMF IS 170 442 00:16:19,323 --> 00:16:20,424 MILLI VOLTS RIGHT NOW BECAUSE 443 00:16:20,424 --> 00:16:21,692 THEY'RE SPINNING AT A REGULAR 444 00:16:21,692 --> 00:16:23,327 NORMAL RATE AND NOW THEY--WITHIN 445 00:16:23,327 --> 00:16:25,396 A FEW SECONDS OF EACH OTHER, 446 00:16:25,396 --> 00:16:27,898 THEY BOTH STOP AND THEN IF 447 00:16:27,898 --> 00:16:29,767 ANOTHER 30 OR 40-SECONDS THEY 448 00:16:29,767 --> 00:16:31,902 ARE GOING TO LYSE, AND 25 449 00:16:31,902 --> 00:16:33,671 MINUTES BEFORE WE TARTED THIS 450 00:16:33,671 --> 00:16:36,374 REALTIME VIDEO, WE EDIT IT OR 451 00:16:36,374 --> 00:16:38,175 INDUCE THE GENES SO WHAT WE SAW 452 00:16:38,175 --> 00:16:39,310 WAS ANOTHER REPRESENTATION OF 453 00:16:39,310 --> 00:16:40,711 THE SAME TIMING EVENT BEFORE 454 00:16:40,711 --> 00:16:42,613 WHERE AFTER A LONG PERIOD OF 455 00:16:42,613 --> 00:16:45,549 TIME IN THE ORDER OF MINUTES, 25 456 00:16:45,549 --> 00:16:47,852 MINUTES PERKAY, IN THIS CASE 457 00:16:47,852 --> 00:16:50,354 THERE WAS A PLAZ MIDS MID, AND 458 00:16:50,354 --> 00:16:52,223 SUDDENLY INTED OF TITRATING IT 459 00:16:52,223 --> 00:16:53,524 OUT SLOWLY, IT ALL HAPPENED AT 460 00:16:53,524 --> 00:16:54,125 ONCE. 461 00:16:54,125 --> 00:16:57,895 SO WE CALLED THIS PAPER, HOLIN'S 462 00:16:57,895 --> 00:16:59,497 KILL WITHOUT WARNING SO THIS WAS 463 00:16:59,497 --> 00:17:01,365 A REPUTATION OF THIS PUKE OUT 464 00:17:01,365 --> 00:17:11,909 MODEL EVERYBODY WAS SUGGESTING. 465 00:17:14,879 --> 00:17:15,079 OKAY. 466 00:17:15,079 --> 00:17:16,814 THIS FTION AN ACCIDENTAL 467 00:17:16,814 --> 00:17:23,354 EXPERIMENT DONE BIAN JELLICCA, 468 00:17:23,354 --> 00:17:25,423 SO BY ELIMINATING THAT YOU COULD 469 00:17:25,423 --> 00:17:26,791 PUT CYSTINES WHEREVER YOU WANT 470 00:17:26,791 --> 00:17:29,093 AND THE IDEA WAS TO USE THESE 471 00:17:29,093 --> 00:17:31,462 CYSTINES TO VARIOUS TYPES OF 472 00:17:31,462 --> 00:17:33,364 BIOCHEMICAL REACTIONS, INCLUDING 473 00:17:33,364 --> 00:17:34,698 DIMERIZATION,OXIDATION, SO ON SO 474 00:17:34,698 --> 00:17:37,201 SHE WEPT THROUGH AND PUT SINGLE 475 00:17:37,201 --> 00:17:39,737 CIS RESIDUES ISSUES THESE ARE 476 00:17:39,737 --> 00:17:40,438 ALL MISSED CHANGES. 477 00:17:40,438 --> 00:17:41,539 SO WE NOTICED THAT THE COLOR 478 00:17:41,539 --> 00:17:43,174 COATED MUSEUM TAIGS IN THE 479 00:17:43,174 --> 00:17:44,775 SECOND AND THIRD TRANSMEMBRANE 480 00:17:44,775 --> 00:17:46,744 DOMAIN THAT FOR EVERY SINGLE 481 00:17:46,744 --> 00:17:48,712 INITANCE CHANGED CAUSED A TIMING 482 00:17:48,712 --> 00:17:51,482 CHANGE IN LYSIS, EVERY SINGLE 1, 483 00:17:51,482 --> 00:17:52,716 SOMETIMES THEY WERE EARLIER, 484 00:17:52,716 --> 00:17:53,984 SOMETIMES THEY WERE LATER AND WE 485 00:17:53,984 --> 00:17:55,853 CALL THIS THE RAINBOW, BECAUSE 486 00:17:55,853 --> 00:17:58,022 THIS GOT US THINKING ABOUT HOW 487 00:17:58,022 --> 00:17:59,090 THIS TIMING CLOCK COULD WORK 488 00:17:59,090 --> 00:18:01,525 EMPLOY SO THAT MEANS THAT--AND 489 00:18:01,525 --> 00:18:04,862 THEN SUPER IMPOSED ON THIS, IF 490 00:18:04,862 --> 00:18:07,765 YOU--THIS WAS DONE BY INDUCING A 491 00:18:07,765 --> 00:18:09,700 LAMDA PHAGE IN TRANSTO THIS PLAZ 492 00:18:09,700 --> 00:18:09,967 MIDS MID. 493 00:18:09,967 --> 00:18:11,402 SO IF YOU COUNT THE PARTICLES 494 00:18:11,402 --> 00:18:13,437 THAT WERE BEING GENERATED AND 495 00:18:13,437 --> 00:18:14,872 REMEMBER, THE PARTICLES ARE 496 00:18:14,872 --> 00:18:16,040 BEING ASSEMBLED ALL THE TIME, 497 00:18:16,040 --> 00:18:19,310 THIS IS GOING ON, AND NORMALLY 498 00:18:19,310 --> 00:18:22,446 AT ABOUT 50 MINUES YOU WOULD 499 00:18:22,446 --> 00:18:25,616 HAVE A HUNDRED OR SO, BUT IF YOU 500 00:18:25,616 --> 00:18:29,320 GO TO 3 HOURS, YOU WOULD 600 SO 501 00:18:29,320 --> 00:18:32,289 IT DOES RAISE THE QUESTION, WHY 502 00:18:32,289 --> 00:18:34,358 DOES LAMDA QUIT AT A HUNDRED, IS 503 00:18:34,358 --> 00:18:35,693 THAT GOOD ENOUGH OR WHAT'S THE 504 00:18:35,693 --> 00:18:36,627 RATIONAL FOR IT AND THE OTHER 505 00:18:36,627 --> 00:18:42,566 THING IS HOW CAN WE EXPLAIN 506 00:18:42,566 --> 00:18:45,236 EVERY SINGLE CHANGE IN 3 507 00:18:45,236 --> 00:18:45,970 DIFFERENT TRANSMEMBRANE DOMAINS 508 00:18:45,970 --> 00:18:47,805 ALTERING THE TIMING AND BY THE 509 00:18:47,805 --> 00:18:49,807 WAY WE CHECKED AND NONE OF THEM 510 00:18:49,807 --> 00:18:50,875 HAVE ANY ACCUMULATION OF THE 511 00:18:50,875 --> 00:18:51,575 PROTEIN. 512 00:18:51,575 --> 00:18:53,777 SO THAT LED TO A PROJECT THAT 513 00:18:53,777 --> 00:18:56,914 REBECCA WHITE RAN AND REBECCA 514 00:18:56,914 --> 00:19:00,017 WAS--SHE'S NOW THE CEO OF A 515 00:19:00,017 --> 00:19:02,620 COMPANY CALLED AVALO, I THINK 516 00:19:02,620 --> 00:19:03,821 IT'S THE THIRD 1 SHE'S RUN BUT 517 00:19:03,821 --> 00:19:06,624 HE MADE THIS INTO A STANDARD 518 00:19:06,624 --> 00:19:09,126 UNDERGRAT GRADUATE LAB, SO EVERY 519 00:19:09,126 --> 00:19:10,161 UNDERGRADUATE THAT WOULD COME TO 520 00:19:10,161 --> 00:19:12,029 THE LAB WOULD GET A MISSENSED 521 00:19:12,029 --> 00:19:13,664 MUSEUM TAIG EMPLOY SO WE HAD 2 522 00:19:13,664 --> 00:19:16,634 ORIGINALS BACK IN THE 70S AND 523 00:19:16,634 --> 00:19:19,069 THESE WERE ALANINE 52 AND 524 00:19:19,069 --> 00:19:19,870 ALA1952 TO GLY. 525 00:19:19,870 --> 00:19:21,539 AND THEY WERE CONSERVATIVE IN 1 526 00:19:21,539 --> 00:19:23,407 SENSE BUT 1'S IMING TO BE A 527 00:19:23,407 --> 00:19:24,241 LARGE OUTFIELD GROUP AND THE 528 00:19:24,241 --> 00:19:26,477 OTHER 1 IS A HYDROGEN ATOM AND 529 00:19:26,477 --> 00:19:28,212 NEITHER 1 OF THEM, SO BOTH OF 530 00:19:28,212 --> 00:19:35,786 THESE MUTATIONS WERE ISOLATED AS 531 00:19:35,786 --> 00:19:36,954 NONPLAQUE FORMERS, SO THAT'S 532 00:19:36,954 --> 00:19:39,456 PROVE THAT IT HAS 2 FUNCTIONS 533 00:19:39,456 --> 00:19:43,494 BECAUSE YOU COULD KNOCK OUT BOTH 534 00:19:43,494 --> 00:19:43,794 OF THEM. 535 00:19:43,794 --> 00:19:47,331 SO IF YOU LOOK AT THIS, SO WHEN 536 00:19:47,331 --> 00:19:49,033 SHE FIRST STARTED OUT WAS TO 537 00:19:49,033 --> 00:19:51,335 HAVE EVERY SINGLE POSSIBLE OTHER 538 00:19:51,335 --> 00:19:56,774 AMINO ACID CHANGE DONE AT ALA52, 539 00:19:56,774 --> 00:19:59,877 AND WE DISCOVER THAD A 4 OR 5 OF 540 00:19:59,877 --> 00:20:01,979 THEM LIESED AT A RIDICULOUSLY 541 00:20:01,979 --> 00:20:03,180 EARLY TIME, SO PHAGES ARE 542 00:20:03,180 --> 00:20:04,615 PRODUCED BY ANY OF THESE, AND 543 00:20:04,615 --> 00:20:07,218 THAT'S WHY YOU DON'T GET ANY 544 00:20:07,218 --> 00:20:08,319 PLAQUES BUT YOU ALSO HAVE A 545 00:20:08,319 --> 00:20:11,355 NUMBER THAT WAS SLOWER AND IF 546 00:20:11,355 --> 00:20:12,990 YOU PUT, YOU KNOW CHARGE 547 00:20:12,990 --> 00:20:14,325 RESIDUES IN THIS THE MIDDLE OF 548 00:20:14,325 --> 00:20:15,593 THE MEMBRANE, YOU WILL KILL THE 549 00:20:15,593 --> 00:20:16,560 MEMBRANE SO THAT WASN'T 550 00:20:16,560 --> 00:20:20,464 SURPRISING THAT A LOT OF 551 00:20:20,464 --> 00:20:22,266 PROTEINS WERE ENACTIVATED SO 552 00:20:22,266 --> 00:20:24,034 THEN SHE EXTEND THAD TO ALL 553 00:20:24,034 --> 00:20:32,977 OTHER, TO THE 3 RESIDUES NEXT TO 554 00:20:32,977 --> 00:20:34,712 THAT, 49, 50, 51 AND 52. 555 00:20:34,712 --> 00:20:38,549 AND BOTTOM LINE IS THAT IT'S THE 556 00:20:38,549 --> 00:20:42,319 SAME FOR 4950 AND 52, BUT IN MED 557 00:20:42,319 --> 00:20:45,623 50, IT WAS SILENT BUT WE GOT 558 00:20:45,623 --> 00:20:50,761 VERY LARGE NUMBER OF EARLY LYSIS 559 00:20:50,761 --> 00:20:51,028 MUTANTS. 560 00:20:51,028 --> 00:20:53,697 SO TAKE HOME NOTES IS THAT LAMDA 561 00:20:53,697 --> 00:20:55,399 COULD EXPLORE TIME SPACE, IF IT 562 00:20:55,399 --> 00:20:57,835 HAS A REASON TO EXPLORE LYSIS 563 00:20:57,835 --> 00:21:00,271 TIME SPACE, IT CAN DO THAT BY 564 00:21:00,271 --> 00:21:02,673 SINGLE CHANGES AND A SINGLE 565 00:21:02,673 --> 00:21:03,874 CONSERVATIVE CHANGE CAN 566 00:21:03,874 --> 00:21:05,175 DRAMATICALLY AFFAIRS TEAM 567 00:21:05,175 --> 00:21:13,050 LEADERRER TIME IN EITHER EARLIER 568 00:21:13,050 --> 00:21:13,517 OR LATER. 569 00:21:13,517 --> 00:21:16,620 OKAY, SO HERE COMES THE FIRST 570 00:21:16,620 --> 00:21:17,154 MODEL. 571 00:21:17,154 --> 00:21:20,758 SO THE MODEL FOR HOLIN'S WORK IS 572 00:21:20,758 --> 00:21:22,226 THAT THE INNER MEMBRANES, I'M 573 00:21:22,226 --> 00:21:23,360 NOT HOEING YOU ANYTHING ABOUT 574 00:21:23,360 --> 00:21:25,529 THE PHAGE. 575 00:21:25,529 --> 00:21:28,399 PHAGE PARTICLES AS LONG AS 576 00:21:28,399 --> 00:21:29,800 THERE'S LYSIS PROTEINS BEING 577 00:21:29,800 --> 00:21:32,870 PROTUSSED THAT'S ALSO THE MORPH 578 00:21:32,870 --> 00:21:34,171 O GENERIC PARTICLE FOR PHAGE 579 00:21:34,171 --> 00:21:35,005 PRODUCED BUT WE'RE NOT SHOWING 580 00:21:35,005 --> 00:21:39,710 THAT IN THE CARTOON, SO BLUE IS 581 00:21:39,710 --> 00:21:41,111 THE WHOLE, LYSOSIEM IN THE GREEN 582 00:21:41,111 --> 00:21:43,881 AND THEY REACH WHAT WE CALL A 583 00:21:43,881 --> 00:21:44,782 CRITICAL CONCENTRATION. 584 00:21:44,782 --> 00:21:48,285 THIS IS THE MODEL ANYWAY. 585 00:21:48,285 --> 00:21:50,154 AND THEN AT THAT CRITICAL 586 00:21:50,154 --> 00:21:51,021 CONCENTRATION, THE PROTEINS 587 00:21:51,021 --> 00:21:54,158 AGGREGATE INTO WHAT WE CALL A 588 00:21:54,158 --> 00:21:57,594 RAFT AND THEN SOMEHOW THAT RAFT 589 00:21:57,594 --> 00:21:59,430 IS CONVERTED INTO HOLES AND 590 00:21:59,430 --> 00:22:04,468 THESE HOLES ARE UNFORTUNATELY 591 00:22:04,468 --> 00:22:05,269 CALLED S-HOLES. 592 00:22:05,269 --> 00:22:05,536 [LAUGHTER] 593 00:22:05,536 --> 00:22:07,905 IT NEVER FAILS TO--ANYWAY, THESE 594 00:22:07,905 --> 00:22:11,975 ARE MIRROR IMAGE CROHN SCALE, 595 00:22:11,975 --> 00:22:14,378 THE LARGEST HOLES FORMED IN 596 00:22:14,378 --> 00:22:16,880 BIOLOGY BY NORMAL MEANS. 597 00:22:16,880 --> 00:22:20,084 AND SO, THESE GIANT HOLES ALLOW 598 00:22:20,084 --> 00:22:21,852 THE LYSOSIEM TO ESCAPE AND 599 00:22:21,852 --> 00:22:22,920 ATTACK THE CELL WALL AND OF 600 00:22:22,920 --> 00:22:27,658 COURSE WE ALREADY SAW THAT LYSIS 601 00:22:27,658 --> 00:22:29,460 IS FUNDAMENTALLY A PUNKITATIVE 602 00:22:29,460 --> 00:22:31,195 END AND THAT'S REFINED BY WHERE 603 00:22:31,195 --> 00:22:36,400 THE HOLE IS FORMED. 604 00:22:36,400 --> 00:22:37,968 OKAY, SO WERE, THE UNIVERSAL 605 00:22:37,968 --> 00:22:40,037 PHENOTYPE IS THE TRIGGERING ALSO 606 00:22:40,037 --> 00:22:41,338 HAS TO BE EXPLAINED SO WHAT WE 607 00:22:41,338 --> 00:22:44,007 ARGUE IS THAT IN THE--IF YOU HAD 608 00:22:44,007 --> 00:22:44,975 SIGH NIGHED AT THIS POINT 609 00:22:44,975 --> 00:22:46,009 INSTEAD OF GOING THROUGH THE 610 00:22:46,009 --> 00:22:47,544 WAITING FOR THE CRITICAL 611 00:22:47,544 --> 00:22:49,046 CONCENTRATION BEFORE YOU FORM 612 00:22:49,046 --> 00:22:51,949 THE RAF, THE ADDITION OF CYANIDE 613 00:22:51,949 --> 00:22:54,718 INITANTLY ARK LOWS WHATEVER THE 614 00:22:54,718 --> 00:22:56,587 LEVEL OF HOLIN IS, TO FORM, AND 615 00:22:56,587 --> 00:22:57,888 IT TURNS OUT IF 1 SIDE 616 00:22:57,888 --> 00:22:59,523 PREDECLARATION OF THAT IS THAT 617 00:22:59,523 --> 00:23:01,592 THE HOLE SHOULD BE SMALLER 618 00:23:01,592 --> 00:23:03,193 BECAUSE THERE'S LESS LYSISSED 619 00:23:03,193 --> 00:23:04,228 PROTEIN AND YOU'LL SEE THAT IS 620 00:23:04,228 --> 00:23:05,829 THE CASE, SO THE FIRST THING YOU 621 00:23:05,829 --> 00:23:07,931 WANT TO SHOW IS TRUE, THAT IT'S 622 00:23:07,931 --> 00:23:16,006 ACTUALLY MOBILE AND SO, IN MORE 623 00:23:16,006 --> 00:23:16,807 CLARRABRAITORS FROM 624 00:23:16,807 --> 00:23:17,808 [INDISCERNIBLE] LAB, THESE ARE 625 00:23:17,808 --> 00:23:21,779 EXPERIMENTS, SO IF YOU TURN ON 626 00:23:21,779 --> 00:23:22,980 THE HOLIN GFP FUSION WHICH WORKS 627 00:23:22,980 --> 00:23:29,052 PRETTY WELL AND IF YOU USE A52 628 00:23:29,052 --> 00:23:31,288 IT JUST STAYS MOBILE SO IT 629 00:23:31,288 --> 00:23:34,324 IMMEDIATELY FOLLOWS OR BECOMES 630 00:23:34,324 --> 00:23:39,563 FLUORESCENT AND IF YOU DO THE 631 00:23:39,563 --> 00:23:42,800 BLEACHING IN THE S105 GFP BEFORE 632 00:23:42,800 --> 00:23:44,401 TRIGGERING, YOU GET REPOPULATION 633 00:23:44,401 --> 00:23:45,602 OF FLUORESCENCE BUT AFTER 634 00:23:45,602 --> 00:23:48,772 TRIGGERING IT'S NO LONGER, IT 635 00:23:48,772 --> 00:23:50,107 DOESN'T RECOVER AS YOU CAN SEE. 636 00:23:50,107 --> 00:23:53,477 THIS THING'S NOT WORKING NOW. 637 00:23:53,477 --> 00:23:53,710 OH YEAH. 638 00:23:53,710 --> 00:23:55,679 SO S105 AND MOBILE AND BEFORE 639 00:23:55,679 --> 00:23:58,849 TRIGGERING AND IMMOBILE AFTER 640 00:23:58,849 --> 00:23:59,183 TRIGGERING. 641 00:23:59,183 --> 00:24:00,517 THIS WAS PROBABLY LE THE HARDEST 642 00:24:00,517 --> 00:24:03,987 EXPERIMENT WE EVER DID, THIS IS 643 00:24:03,987 --> 00:24:07,257 S-BIOCHEMISTRY, I WON'T ANY INTO 644 00:24:07,257 --> 00:24:11,361 THE DESPERATION MODE TO PURIFY 645 00:24:11,361 --> 00:24:15,065 HOLEINS, TURNS OUT YOU CAN GET 646 00:24:15,065 --> 00:24:17,434 HALF A GRAM PER LITER BUT WE 647 00:24:17,434 --> 00:24:20,103 WERE ABLE TO PURIFY THE LAMDA 648 00:24:20,103 --> 00:24:22,172 HOLIN, AND I HAVE TO TELL YOU WE 649 00:24:22,172 --> 00:24:23,240 HAD TO EXPLORE TREAN DIFFERENT 650 00:24:23,240 --> 00:24:24,908 POSITIONS TO PUT THAT TAG IN 651 00:24:24,908 --> 00:24:27,044 BEFORE WE FOUND 1 THAT WOULD 652 00:24:27,044 --> 00:24:27,244 WORK. 653 00:24:27,244 --> 00:24:29,513 SO THIS WAS WORKED ON MY JOHN 654 00:24:29,513 --> 00:24:30,747 AND DAVE SMITH. 655 00:24:30,747 --> 00:24:33,817 O YOU BASICALLY PURIFY IT, DUMP 656 00:24:33,817 --> 00:24:35,919 IT INTO A KOTROAP WHERE IT'S 657 00:24:35,919 --> 00:24:38,322 TOTALLY UNFOLDED AND THEN YOU 658 00:24:38,322 --> 00:24:41,525 DILUTE THAT DOWN INTO SUSPENSION 659 00:24:41,525 --> 00:24:43,594 OF LYSOSOMES THAT HAVE 660 00:24:43,594 --> 00:24:44,294 FLUORESCENT CALCIUM IN THEM AND 661 00:24:44,294 --> 00:24:45,729 THEN WHAT YOU SLEEP APNEA AND 662 00:24:45,729 --> 00:24:47,798 OBESITYY IS RELEASE THE 663 00:24:47,798 --> 00:24:50,834 WILD-TYPE RELEASES THE 664 00:24:50,834 --> 00:24:51,668 FLUORESCENCE, THE MUTANT DOESN'T 665 00:24:51,668 --> 00:24:52,803 AT EVEN TWICE THE CONITRATION 666 00:24:52,803 --> 00:24:54,771 AND EVEN MORE IMPORTANTLY 667 00:24:54,771 --> 00:24:57,441 PURIFIED MATERIAL FROM A 668 00:24:57,441 --> 00:24:59,977 TS-ALLELE WHICH IS ALOE 19 TO 3 669 00:24:59,977 --> 00:25:01,278 A19, WORKS AT THE HIGH 670 00:25:01,278 --> 00:25:03,013 TEMPERATURE, NOT THE LOW 671 00:25:03,013 --> 00:25:03,947 TEMPERATURE WHICH FORMERLY 672 00:25:03,947 --> 00:25:06,250 EXCLOUDS THE POSSIBILITY OF SOME 673 00:25:06,250 --> 00:25:07,951 CONTAMINANT OF SOME PREPARATION 674 00:25:07,951 --> 00:25:09,653 THAT WAS CAUSING FORMULATION SO 675 00:25:09,653 --> 00:25:12,322 IT DOES WHAT IT'S SUPPOSED TO DO 676 00:25:12,322 --> 00:25:13,524 AND CAN PERMIALIZE THE MEMBRANES 677 00:25:13,524 --> 00:25:20,597 IN AN ARK LEGAL SPECIFIC MANNER 678 00:25:20,597 --> 00:25:26,436 I THINK THIS IS SO JUST TO 679 00:25:26,436 --> 00:25:29,239 REMIND YOU WHAT HAPPENSA THE 680 00:25:29,239 --> 00:25:29,940 MILLI SECURITIZATION. LEVEL, 681 00:25:29,940 --> 00:25:36,880 THERE'S GOING TO BE A EXPULSION 682 00:25:36,880 --> 00:25:37,548 OF THE INNER--CYTOPLASMIC 683 00:25:37,548 --> 00:25:39,416 MATERIAL FOR THE LOCAL BLOW OUT. 684 00:25:39,416 --> 00:25:42,519 OKAY, SO OUR NEXT GOAL WAS TO 685 00:25:42,519 --> 00:25:44,588 VISUALIZE THE S-HOLE AND IT 686 00:25:44,588 --> 00:25:45,989 SHOULD BE BIG ENOUGH, WELL THIS 687 00:25:45,989 --> 00:25:48,325 IS HOW WE LEARN IF IT'S MICRON 688 00:25:48,325 --> 00:25:48,625 SCALE. 689 00:25:48,625 --> 00:25:51,194 THIS WAS A COLLABORATION WITH 690 00:25:51,194 --> 00:25:54,298 ANDRE'S LAB AT [INDISCERNIBLE], 691 00:25:54,298 --> 00:25:55,299 AND [INDISCERNIBLE] DID THE WORK 692 00:25:55,299 --> 00:25:57,501 AND SO BASICALLY YOU'RE LOOKING 693 00:25:57,501 --> 00:25:59,036 AT INTERRUPTIONS IN THE MEMBRANE 694 00:25:59,036 --> 00:26:03,607 OF THE E.COLI AND THE AVERAGE 695 00:26:03,607 --> 00:26:05,175 AND THE AVERAGE SIZE OF THESE, 696 00:26:05,175 --> 00:26:06,476 ACTUALLY LET ME SHOW YOU THE 697 00:26:06,476 --> 00:26:09,246 NEXT SLIDE, WHICH IS--YEAH, THIS 698 00:26:09,246 --> 00:26:15,652 IS ACTUALLY A TOMOGRAM. 699 00:26:15,652 --> 00:26:17,054 IT TOOK US A YEAR TO DO THIS 700 00:26:17,054 --> 00:26:19,189 EXPERIMENT AND THEN A REVIEWER 701 00:26:19,189 --> 00:26:20,223 RYED ANOTHER 1 AND IT TOOK 702 00:26:20,223 --> 00:26:21,491 ANOTHER 1 TO DO THE EXPERIMENT 703 00:26:21,491 --> 00:26:25,662 BUT THIS WAS ABOUT A HUNDRED 704 00:26:25,662 --> 00:26:27,397 MICRON HOLD, ON AVERAGE THERE'S 705 00:26:27,397 --> 00:26:30,334 1 TO 3 PER CELL AND ON AVERAGE 706 00:26:30,334 --> 00:26:31,368 THEY ARE 350-MICRONS. 707 00:26:31,368 --> 00:26:33,070 SO EVERYBODY WANTED TO SEE THE 708 00:26:33,070 --> 00:26:34,137 HOLE AND WE DID IT. 709 00:26:34,137 --> 00:26:35,706 SO THEN THE NEXT THING WAS, 710 00:26:35,706 --> 00:26:37,174 WHAT'S THE STRUCTURE OF THE 711 00:26:37,174 --> 00:26:40,210 HOLE, AND SO WE KNEW THAT THE 712 00:26:40,210 --> 00:26:42,512 S-PROTEIN STARTED OW AS A MOBILE 713 00:26:42,512 --> 00:26:43,447 PROBABLY DISCIPLINARY MERRIC 714 00:26:43,447 --> 00:26:44,648 FORM THAT FOLDED AROUND THE 715 00:26:44,648 --> 00:26:47,584 MEMBRANE AND ENDS UP IN AN 716 00:26:47,584 --> 00:26:48,752 IMMOBILE LARGE MICRON SCALE HOLE 717 00:26:48,752 --> 00:26:49,953 BUT WHAT'S THE ACTUAL STRUCTURE 718 00:26:49,953 --> 00:26:52,356 AND WHAT'S FACING WHAT EMPLOY SO 719 00:26:52,356 --> 00:26:55,459 WHAT WE WANTED TO DO WAS TO FIND 720 00:26:55,459 --> 00:26:59,863 OUT IN THE S-MOLECULE, REMEMBER 721 00:26:59,863 --> 00:27:03,533 I TOLD YOU ABOUT ANGELICA'S WORK 722 00:27:03,533 --> 00:27:04,434 FOR PUTTING CYSTINES EVERYWHERE, 723 00:27:04,434 --> 00:27:06,103 CAN YOU SEE ALL THE CYSTINES WE 724 00:27:06,103 --> 00:27:08,171 USED AND THEN YOU TREAT THIS 725 00:27:08,171 --> 00:27:11,541 WITH A REAGENT AND IT BLOCKS AND 726 00:27:11,541 --> 00:27:13,510 IF THE REAGENT MODIFIED PROTEIN 727 00:27:13,510 --> 00:27:14,678 IT'S IN THE SOLUBLE PHASE AND 728 00:27:14,678 --> 00:27:16,647 YOU CAN DETECH THAT BY WESTERN 729 00:27:16,647 --> 00:27:19,516 BLOTS BECAUSE YOU PAY FOR THAT 730 00:27:19,516 --> 00:27:20,584 AFTERWARDS, THE BOTTOM LINE WAS 731 00:27:20,584 --> 00:27:22,986 THAT THERE WERE HALF OF TMD 1 732 00:27:22,986 --> 00:27:25,656 AND 3 THAT WERE EXPOSED IN THE 733 00:27:25,656 --> 00:27:30,394 HOLE, WERE EXPOSED TO THE 734 00:27:30,394 --> 00:27:30,961 AQUESTIONNAIRESUS SOLUTION 735 00:27:30,961 --> 00:27:34,431 WHEREAS IN THE MEMBRANE WHEREAS 736 00:27:34,431 --> 00:27:35,832 TMD2 IN THE MIDDLE WAS EXPOSED 737 00:27:35,832 --> 00:27:40,170 SO THAT,A LOWED US AND KENNETH 738 00:27:40,170 --> 00:27:43,607 TOW WHO NOW RUNS [INDISCERNIBLE] 739 00:27:43,607 --> 00:27:47,210 WHO DID THIS WORK AND THIS LED 740 00:27:47,210 --> 00:27:51,448 TO THIS MODEL WHERE HYDROPHILIC 741 00:27:51,448 --> 00:27:53,750 PHASES OF TMD'S 1 AND 3 IN THE 742 00:27:53,750 --> 00:27:55,185 DIMER AND I'M NOT SHOWING YOU 743 00:27:55,185 --> 00:27:57,721 THE EVIDENCE, IT'S A DIMER IT'S 744 00:27:57,721 --> 00:27:59,456 VERY SIMPLE CROSS LINKING 745 00:27:59,456 --> 00:28:02,626 EXPERIMENTS, SO, BASED ON WHAT 746 00:28:02,626 --> 00:28:04,294 WE JUST SAW, THE SYSTEM WITH THE 747 00:28:04,294 --> 00:28:05,996 NOTION THAT THE HYDROPHILIC 748 00:28:05,996 --> 00:28:08,365 PHASES AND MULTIPLE DIMER ARE 749 00:28:08,365 --> 00:28:09,533 FACING EACH OTHER SEQUESTER ON 750 00:28:09,533 --> 00:28:11,068 THE MEMBRANE, SO WE'RE LOOKING 751 00:28:11,068 --> 00:28:12,169 DOWN, SO THIS,A COUNTS FOR THE 752 00:28:12,169 --> 00:28:16,106 ABILITY OF THE PROTEIN TO 753 00:28:16,106 --> 00:28:19,576 FREELYY MIGRATE BECAUSE IT'S 754 00:28:19,576 --> 00:28:21,078 NOT--ITS HYDROPHILIC PHASES ARE 755 00:28:21,078 --> 00:28:21,912 NOT FACING ANY OTHER SIDE. 756 00:28:21,912 --> 00:28:24,648 SO IN THE LUMEN, IN THE FINAL 757 00:28:24,648 --> 00:28:28,685 TRUCTURE AFTER COLLAPSE, THIS IS 758 00:28:28,685 --> 00:28:30,087 THE MODEL WE SAY BECAUSE WE'RE 759 00:28:30,087 --> 00:28:31,521 SHOWING HALF THE FACE OF 1 AND 3 760 00:28:31,521 --> 00:28:38,195 ARE ACCESSIBLE TO THE SOLUBLE 761 00:28:38,195 --> 00:28:38,428 REAGENT. 762 00:28:38,428 --> 00:28:41,565 AND, BY THE WAY, WE MEASURED THE 763 00:28:41,565 --> 00:28:43,533 AMOUNT OF S-PROTEIN BY 764 00:28:43,533 --> 00:28:44,468 QUANTITATIVE WESTERN BLOT AND 765 00:28:44,468 --> 00:28:47,504 ALSO BY SOMETHING MEASURING THE 766 00:28:47,504 --> 00:28:49,973 PERIMETER OF THE HALL ANDS IF 767 00:28:49,973 --> 00:28:54,044 YOU FILLED ON THE FACT THAT EACH 768 00:28:54,044 --> 00:28:56,580 1 OF THESE ALPHA HELIXES IS 769 00:28:56,580 --> 00:28:58,782 ABOUT 1 NANOMETER IN DIAMETER 770 00:28:58,782 --> 00:29:00,984 THERE'S A CONSISTENT NUMBER OF 771 00:29:00,984 --> 00:29:02,385 S105 MOLECULES IN THE TOTAL 772 00:29:02,385 --> 00:29:12,796 PERIMETER OF THE HOLES. 773 00:29:19,269 --> 00:29:20,437 HERE IS THE GRAPED MODEL HERE, 774 00:29:20,437 --> 00:29:22,839 SO WE HAVE A MUTANT THAT DOESN'T 775 00:29:22,839 --> 00:29:23,940 DIMERIZE SO WE SUSPECT 776 00:29:23,940 --> 00:29:25,509 THAT--WELL WE DON'T KNOW WHAT 777 00:29:25,509 --> 00:29:30,013 CONTROLS THE DIMERIZATION, BUT 778 00:29:30,013 --> 00:29:32,115 THE DIMER IS THE MOLECULE THAT 779 00:29:32,115 --> 00:29:32,849 UNDERGOES THE TIMING BECAUSE 780 00:29:32,849 --> 00:29:35,719 IT'S NOT UNTIL THE DIMER REACHES 781 00:29:35,719 --> 00:29:37,487 CRITICAL CONCENTRATION THAT IT 782 00:29:37,487 --> 00:29:39,456 SUDDENLY AGGREGATES TO FORMER 783 00:29:39,456 --> 00:29:41,858 RAF AND WE'RE ARGUING THAT THE 784 00:29:41,858 --> 00:29:43,393 TOTAL POPULATION OF S-MOLLULES 785 00:29:43,393 --> 00:29:45,028 IN RAF IS ABOUT 5000 AND IT'S 786 00:29:45,028 --> 00:29:47,597 THE EXISTENCE OF THE RAF THAT 787 00:29:47,597 --> 00:29:51,268 CAUSES THE NEXT PHASE AND HERE'S 788 00:29:51,268 --> 00:29:52,769 WHERE--I WILL BEG YOUR 789 00:29:52,769 --> 00:29:54,304 INDULGENCE SO WHAT WE THINK IS 790 00:29:54,304 --> 00:29:56,239 HAPPENING IS, YOU HAVE A SHORT 791 00:29:56,239 --> 00:29:59,876 CIRCUIT BECAUSE YOU HAVE A LARGE 792 00:29:59,876 --> 00:30:02,946 RELATIVELY LARGE SURFACE OF 793 00:30:02,946 --> 00:30:04,881 LIPID DEPLETED ASHES JACENT 794 00:30:04,881 --> 00:30:06,416 PROTEIN MOLECULES WHICH ARE 795 00:30:06,416 --> 00:30:07,517 FEATURED BY TRANSMEMBRANE 796 00:30:07,517 --> 00:30:11,721 DOMAINS EACH 1 WITH AN ACTIVE 797 00:30:11,721 --> 00:30:12,556 POLAR, SEVERAL ACTIVE POLAR 798 00:30:12,556 --> 00:30:15,091 UNITS SO WE THINK THERE'S A 799 00:30:15,091 --> 00:30:16,593 SHORT CIRCUIT LOCAL DEPOLAR 800 00:30:16,593 --> 00:30:18,361 EYATION IN THE RAF, AND NOW YOU 801 00:30:18,361 --> 00:30:21,531 ARE BACK TO OUR ORIGINAL IDEA IF 802 00:30:21,531 --> 00:30:23,533 YOU DEPOLARIZE THE MEMBRANE THAT 803 00:30:23,533 --> 00:30:26,770 CAUSES THE SECONDARY, TERTIARY 804 00:30:26,770 --> 00:30:28,305 AND QUANDARY CHANGE THAT STARTS 805 00:30:28,305 --> 00:30:29,039 THE FORMATION OF THE HOLE. 806 00:30:29,039 --> 00:30:31,274 SO IT'S THE FORMATION OF RAF 807 00:30:31,274 --> 00:30:33,777 THAT LEADS TO THE SMALL HOLES 808 00:30:33,777 --> 00:30:35,212 AND RAPIDLY EXPAND AND FEET 809 00:30:35,212 --> 00:30:38,181 FORWARD MECHANISM TO THE LARGE 810 00:30:38,181 --> 00:30:39,816 SCALE 300-MICRON HOLES. 811 00:30:39,816 --> 00:30:46,089 SO THAT'S THE MODEL. 812 00:30:46,089 --> 00:30:48,491 AND THEN EITHER PREMATURE LYSIS 813 00:30:48,491 --> 00:30:50,360 OR A LIAISONSIS MUTEITANT THAT 814 00:30:50,360 --> 00:30:51,761 HAD AN EARLIER TRIGGER CHICAGO 815 00:30:51,761 --> 00:30:53,063 WAS EASY TO CREATE. 816 00:30:53,063 --> 00:30:55,031 SIMPLY CAUSES THE CRITICAL 817 00:30:55,031 --> 00:30:57,067 CONCENTRATION TO BE LOWER SO YOU 818 00:30:57,067 --> 00:30:59,436 END UP WITH A SMALLER HOLE WITH 819 00:30:59,436 --> 00:31:00,937 FEWER S-PROTEINS ON THE 820 00:31:00,937 --> 00:31:01,671 PERIMETER APPROXIMATE WE'VE 821 00:31:01,671 --> 00:31:02,172 PUBLISHED THAT. 822 00:31:02,172 --> 00:31:04,507 THAT'S EXACT LYE WHAT HAPPENS TO 823 00:31:04,507 --> 00:31:07,510 THE IMMUNE DISORDER IN PREMATURE 824 00:31:07,510 --> 00:31:10,347 TRIGGERING, YOU REDUCE THE SIZE 825 00:31:10,347 --> 00:31:11,381 OF THE HOLES. 826 00:31:11,381 --> 00:31:17,320 OKAY SO WHAT DO WE MEAN BY 827 00:31:17,320 --> 00:31:17,854 CRITICAL CONCENTRATION. 828 00:31:17,854 --> 00:31:19,456 I DID NOT MAKE THISSA WORD UP. 829 00:31:19,456 --> 00:31:21,424 SO THIS IS WORK DONE BY MARK 830 00:31:21,424 --> 00:31:22,692 KREB'S LAB QUITE A LONG TIME 831 00:31:22,692 --> 00:31:23,793 AGO, HE WAS FROM [INDISCERNIBLE] 832 00:31:23,793 --> 00:31:31,635 LAB AND HE WORKED ON THE P A RVO 833 00:31:31,635 --> 00:31:32,569 VACCINE XTHEY DORPHED VERY EARLY 834 00:31:32,569 --> 00:31:34,804 ON IF YOU PURIFIED IT AND THEN 835 00:31:34,804 --> 00:31:36,873 DILUTED IT INTO A DETERGENT OF 836 00:31:36,873 --> 00:31:41,177 SOME PERCENTAGE THEN IT WOULD 837 00:31:41,177 --> 00:31:42,512 FORM--IS IT FINALLY NOT WORKING 838 00:31:42,512 --> 00:31:52,722 AT ALL NOW. 839 00:31:56,626 --> 00:31:56,793 WHOOPS. 840 00:31:56,793 --> 00:32:00,196 SO THE BOTTOM LINE IS AT 841 00:32:00,196 --> 00:32:02,132 WHATEVER CONCENTRATION YOU MADE 842 00:32:02,132 --> 00:32:03,233 THE PROTEIN, LOW CONCENTRATIONS 843 00:32:03,233 --> 00:32:05,268 IT WOULD ALL BE IN MONITOR MER, 844 00:32:05,268 --> 00:32:08,271 BUT AS YOU WENT TO HIGHER AND 845 00:32:08,271 --> 00:32:10,240 HIGHER CONCENTRATIONS THESE ARE 846 00:32:10,240 --> 00:32:11,308 SUCROSE GRADIENTS, YOU GET MORE 847 00:32:11,308 --> 00:32:12,909 AND MORE IN THE LATTICE FORM SO 848 00:32:12,909 --> 00:32:14,711 IF YOU PLOT THE AMOUNT IN THE 849 00:32:14,711 --> 00:32:16,346 LATTICE VERSUS THE 850 00:32:16,346 --> 00:32:17,480 CONCENTRATION, CAN YOU OCCUR 851 00:32:17,480 --> 00:32:18,982 LIKE THIS, IS WHERE IT 852 00:32:18,982 --> 00:32:20,617 INTERSECTS THE LINE IS THE 853 00:32:20,617 --> 00:32:21,451 CRITICAL CONCENTRATION AND SO IT 854 00:32:21,451 --> 00:32:26,790 TURNS OUT THAT THEY HAD A ROUGH 855 00:32:26,790 --> 00:32:28,291 CRYOEM STRUCTURE OF HOW IT WAS 856 00:32:28,291 --> 00:32:29,526 INTERACTING WITH THE UNITS 857 00:32:29,526 --> 00:32:30,760 INTERACTING WITH EACH OTHER AND 858 00:32:30,760 --> 00:32:33,330 THEY DID SOME SATURATED IMMUNO 859 00:32:33,330 --> 00:32:36,800 GENESIS, AND SO NORMALIZED TO 860 00:32:36,800 --> 00:32:38,768 THE WILD-TYPE OF I HIGHLIGHTED 861 00:32:38,768 --> 00:32:41,137 WOTHAT ARE SIMPLE RELATIVELY 862 00:32:41,137 --> 00:32:41,938 CONSERVATIVE CHANGES THAT EITHER 863 00:32:41,938 --> 00:32:43,573 CHANGE THE CRITICAL 864 00:32:43,573 --> 00:32:44,908 CONCENTRATION OF A FOLD LOWER OR 865 00:32:44,908 --> 00:32:48,178 6 FOLD HIGHER WHICH WHEN IT DOES 866 00:32:48,178 --> 00:32:58,521 THAT IT COMPLETELY CHANGES THE 867 00:32:58,521 --> 00:33:03,460 PERCENTAGE THAT CHANGES THE 868 00:33:03,460 --> 00:33:04,194 CRITICAL CONCENTRATION. 869 00:33:04,194 --> 00:33:06,329 SO I HOPE IT VALIDATED IN YOUR 870 00:33:06,329 --> 00:33:08,365 MIND IT'S THE SINGLE REASON WHY 871 00:33:08,365 --> 00:33:09,966 SINGLE AMINO CHANGES IN THE 872 00:33:09,966 --> 00:33:11,701 HOLIN, YOU ARE CHANGING THE 873 00:33:11,701 --> 00:33:13,036 CONCENTRATION OF WHICH IT 874 00:33:13,036 --> 00:33:14,204 SUDDENLY FORMS THE RAFT. 875 00:33:14,204 --> 00:33:17,474 SO FINAL ACT OF THIS IS TO SHOW 876 00:33:17,474 --> 00:33:18,742 THERE'S ACTUALLY BEHAVES LIKE WE 877 00:33:18,742 --> 00:33:22,712 SAY IT DOES SO THESE ARE FGFP 878 00:33:22,712 --> 00:33:24,114 SPRPTS WHERE IT'S ATTACKED WITH 879 00:33:24,114 --> 00:33:27,283 THE LINKER TO THE C-TERMINAL END 880 00:33:27,283 --> 00:33:28,852 AND CYTOPLASMIC DOMAIN OF THE 881 00:33:28,852 --> 00:33:32,956 HOLIN AND THESE ARE JUST NOW 882 00:33:32,956 --> 00:33:35,058 COMING OUT IN M-BIOSO IT'S 883 00:33:35,058 --> 00:33:39,629 RELATIVELY NEW EMPLOY CAN I GET 884 00:33:39,629 --> 00:33:40,263 THIS CLIENT? 885 00:33:40,263 --> 00:33:46,102 SO THE IDEA IS THAT YOU WILL SEE 886 00:33:46,102 --> 00:33:49,672 SUDDENLY WE START WITH DISPERSED 887 00:33:49,672 --> 00:33:51,941 SGFP AND THEN AG TBREIGATE AND 888 00:33:51,941 --> 00:33:53,943 THEY BOTH FOLD BO RAFTS WHICH 889 00:33:53,943 --> 00:33:55,645 THEN DISAPPEAR APPROXIMATEEE 890 00:33:55,645 --> 00:33:56,379 THINK THE DISAPPEARANCE OF THE 891 00:33:56,379 --> 00:33:58,081 RAFT IS WHEN THE ELONGATION 892 00:33:58,081 --> 00:34:02,552 OCCURS AND THEN YOU GET THE 893 00:34:02,552 --> 00:34:07,057 EXPLOSIVE DISRUPTION OF CELL IN 894 00:34:07,057 --> 00:34:08,758 BOTH OF THESE POSITIONS. 895 00:34:08,758 --> 00:34:09,893 SOONER OR LATER. 896 00:34:09,893 --> 00:34:11,895 THIS IS ONLY SECONDS, IT SEEMS 897 00:34:11,895 --> 00:34:22,205 LONGER WHEN I'M LOOKING AT IT. 898 00:34:22,205 --> 00:34:23,239 THERE WE GO. 899 00:34:23,239 --> 00:34:30,246 IT LOOKS PAINFUL ACTUALLY. 900 00:34:30,246 --> 00:34:31,714 SO THERE'S ANOTHER 1 EMPLOY THIS 901 00:34:31,714 --> 00:34:34,317 WAS A LITTLE BIT MORE CONVINCING 902 00:34:34,317 --> 00:34:37,087 BECAUSE IT'S AT THE POLE, BUT 903 00:34:37,087 --> 00:34:39,923 80% OF THE TIME IT'S AT THE 904 00:34:39,923 --> 00:34:40,890 POLE, IN THIS CASE THEY FORMED 905 00:34:40,890 --> 00:34:51,434 AND THEY WERE NOT AT THE POLAR. 906 00:34:59,175 --> 00:35:00,510 AFTER ALL THAT WORK. 907 00:35:00,510 --> 00:35:00,710 SORRY. 908 00:35:00,710 --> 00:35:02,145 WE'RE RUNNING OUT OF TIME SO LET 909 00:35:02,145 --> 00:35:06,049 ME GO ON TO THE NEXT 1. 910 00:35:06,049 --> 00:35:08,418 SO THERE'S JESSE DEVELOPED 911 00:35:08,418 --> 00:35:10,520 ANOTHER ASSAY FOR THE INTEGRITY 912 00:35:10,520 --> 00:35:12,155 OF THE MEMBRANE, OUR MODEL 913 00:35:12,155 --> 00:35:13,923 SPECIFICALLY PREDICS THAT THE 914 00:35:13,923 --> 00:35:16,726 MEMBRANE WILL DEPOLARIZE BEFORE 915 00:35:16,726 --> 00:35:19,896 THE FIRST DEFORMATION OF THE 916 00:35:19,896 --> 00:35:21,498 CELL MORPHOLOGY OCCURS SO HE'S 917 00:35:21,498 --> 00:35:24,934 MAKING USE OF THE FLUORESCENCE 918 00:35:24,934 --> 00:35:26,803 MOLECULE BIOFLAVIN T, WHICH IS 919 00:35:26,803 --> 00:35:28,972 NORMALLY NOT USED BECAUSE IT 920 00:35:28,972 --> 00:35:31,574 BINDS TO SURFACES AND 921 00:35:31,574 --> 00:35:32,308 FLUORESCENCES, IF YOU TREAT 922 00:35:32,308 --> 00:35:34,477 WHOLE CELLS WITH THAT YOU DON'T 923 00:35:34,477 --> 00:35:35,745 SEE FLUORESCENCE UNTIL THE 924 00:35:35,745 --> 00:35:36,946 MISDEMEANOR BRAIN IS COMPROMISED 925 00:35:36,946 --> 00:35:39,149 AND IT TURNS OUT, HOPEFULLY THIS 926 00:35:39,149 --> 00:35:41,117 PARTICULAR MOVIE HAS CRASHED 927 00:35:41,117 --> 00:35:42,085 NUMBERS OF TIMES. 928 00:35:42,085 --> 00:35:43,553 HOPEFULLY IT DOESN'T. 929 00:35:43,553 --> 00:35:45,155 OKAY, SO BIOFLAVIN T IS PRESENT. 930 00:35:45,155 --> 00:35:47,423 VERY NEAR THE END, WITHIN 931 00:35:47,423 --> 00:35:54,264 SECONDS OF TRIGGERING AND WHAT 932 00:35:54,264 --> 00:35:55,899 YOU SEE IS IT'S FAIRLY UNIFORM 933 00:35:55,899 --> 00:36:01,137 IN THE CELL LONG BEFORE THE UDEN 934 00:36:01,137 --> 00:36:04,107 EXPLOSION AT 1 POLE AND WE HAVE 935 00:36:04,107 --> 00:36:06,276 MANY, MANY MOVIES LIKE THIS. 936 00:36:06,276 --> 00:36:09,746 SO WHAT TIME AM I AT NOW? 937 00:36:09,746 --> 00:36:10,246 >> [INDISCERNIBLE] 938 00:36:10,246 --> 00:36:12,916 >> SO I HAVE TO PREVENTIVELY 939 00:36:12,916 --> 00:36:14,450 TAKE A DIVERGENCE BECAUSE AFTER 940 00:36:14,450 --> 00:36:15,718 WE STARRED THIS CRUSADE TO 941 00:36:15,718 --> 00:36:17,420 UNDERSTAND THE LAM TAKEN--THEY 942 00:36:17,420 --> 00:36:18,788 HOLIN, THE MOST IMPORTANT 943 00:36:18,788 --> 00:36:19,822 PROTEIN AND MOST IMPORTANT 944 00:36:19,822 --> 00:36:21,858 ORLGANISM IN THE WORLD. 945 00:36:21,858 --> 00:36:24,494 IT TURNS OUT THAT IF WE HAD 946 00:36:24,494 --> 00:36:25,595 LISTENED TO LAMDA, LAMDA WAS 947 00:36:25,595 --> 00:36:27,163 SAYING ABOUT YOU WAIT, BUT WAIT. 948 00:36:27,163 --> 00:36:29,499 IT TURNS OUT THERE'S 2 949 00:36:29,499 --> 00:36:33,136 FUNCTIONAL TYPES OF HOLINs, WE 950 00:36:33,136 --> 00:36:36,673 CALL THE LAMDA CONONICLE, BUT 951 00:36:36,673 --> 00:36:39,475 THERE ALSO'S PIN HOLINS WHICH 952 00:36:39,475 --> 00:36:42,045 MAKE TINY HOLES, AND LAMDA TOLD 953 00:36:42,045 --> 00:36:44,447 US THIS BECAUSE IT HAS A NEAREST 954 00:36:44,447 --> 00:36:45,648 COULD YOU SAYIN, THIS IS WHAT 955 00:36:45,648 --> 00:36:50,887 YOU GET TO BE 78, I HAVE AN 956 00:36:50,887 --> 00:36:53,323 UNTIED SHOE LACE. 957 00:36:53,323 --> 00:36:55,558 SO THE LAMDA PHAGE IT LOOKS IKE 958 00:36:55,558 --> 00:36:56,726 DENTICAL TO LAMDA BUT WHEN YOU 959 00:36:56,726 --> 00:36:58,795 LOOK AT IT IT'S A HOLEIN AND 960 00:36:58,795 --> 00:37:02,265 ONLY HAS 2 TRANSMEMBRANE DOMAINS 961 00:37:02,265 --> 00:37:04,667 AND THE ENDOLICENSE IS A 962 00:37:04,667 --> 00:37:06,402 LYSOSIEM AND IT HAS WHAT APERS 963 00:37:06,402 --> 00:37:08,404 TO BE A TRANSMEMBRANE DOMAIN OR 964 00:37:08,404 --> 00:37:10,039 SIGNAL SEQUENCE AT THE IMMUNO 965 00:37:10,039 --> 00:37:12,108 TERMINUS, SO THAT WAS A SHOCK. 966 00:37:12,108 --> 00:37:17,513 IF I TRIP AND BREAK MY NECK THEN 967 00:37:17,513 --> 00:37:18,915 I'LL--I WILL NOT GO THROUGH 968 00:37:18,915 --> 00:37:19,949 THIS. 969 00:37:19,949 --> 00:37:21,784 THIS IS 2 SEMINARS, REALLY 970 00:37:21,784 --> 00:37:23,920 AMAZING WORK TO ME BECAUSE WE 971 00:37:23,920 --> 00:37:26,589 DOCUMENTED 2 DIFFERENT TYPES OF 972 00:37:26,589 --> 00:37:27,423 MEMBRANE DYNAMICS, MEMBRANES 973 00:37:27,423 --> 00:37:30,193 YUSMING IN ASK OUT OF MEMBRANES, 974 00:37:30,193 --> 00:37:30,860 TRANSMEMBRANE DOMAINS JUMPING IN 975 00:37:30,860 --> 00:37:32,228 AND OUT OF MEMBRANES SO THE 976 00:37:32,228 --> 00:37:39,402 BOTTOM LINE IS THAT THERE IS 977 00:37:39,402 --> 00:37:41,170 THIS TRANSMEMBRANE COMES OUT AND 978 00:37:41,170 --> 00:37:42,538 THIS TRANSMEMBRANE COMES OUT AND 979 00:37:42,538 --> 00:37:44,073 IT RELEASES AND ACTIVATES THE 980 00:37:44,073 --> 00:37:45,642 ACTIVITY OF THE ENZYME AND THERE 981 00:37:45,642 --> 00:37:47,243 ARE MULTIPLE PAPERS, I ACTUALLY 982 00:37:47,243 --> 00:37:49,245 HAD A REALLY CUTE, MAYBE NOT SO 983 00:37:49,245 --> 00:37:50,413 CUTE COMMENT FROM 1 OF THE 984 00:37:50,413 --> 00:37:52,615 REVIEWERS WHO MAY BE IN THIS 985 00:37:52,615 --> 00:37:54,017 ROOM, SAID THAT YOU REALLY 986 00:37:54,017 --> 00:37:55,652 ADMIRE OUR WORK BUT HE FELT WE 987 00:37:55,652 --> 00:37:58,121 MIGHT HAVE MORE REVIEWERS THAN 988 00:37:58,121 --> 00:37:58,354 READERS. 989 00:37:58,354 --> 00:38:01,457 SO THIS IS A LOT OF VERY HIGH 990 00:38:01,457 --> 00:38:02,825 PROFILE PAPERS, ABOUT A VERY 991 00:38:02,825 --> 00:38:05,161 SMALL THING BUT IT WAS REALLY 992 00:38:05,161 --> 00:38:06,162 HIGH COOL FACTOR IN MY OPINION. 993 00:38:06,162 --> 00:38:10,867 SO I WILL SUMMARIZE HOW THIS 994 00:38:10,867 --> 00:38:11,134 WORKS. 995 00:38:11,134 --> 00:38:12,502 I'VE FLIPPED THROUGH THE WHOLE 996 00:38:12,502 --> 00:38:14,237 THING SO BOTTOM LINE IS, IT'S 997 00:38:14,237 --> 00:38:16,005 THE SAME STORY, JUDGE UOF THE A 998 00:38:16,005 --> 00:38:17,740 SLIGHTLY DIFFERENT, MUSICAL 999 00:38:17,740 --> 00:38:20,610 SCORE, SO, THE HOLIN 1000 00:38:20,610 --> 00:38:21,944 ACCUMULATES, EEIVETTUALLY FOR 1001 00:38:21,944 --> 00:38:23,546 THE PURPOSES RAFTS AT CRITICAL 1002 00:38:23,546 --> 00:38:27,950 CONCENTRATION, AT THAT POINT THE 1003 00:38:27,950 --> 00:38:30,720 ONLY HOLE FORMING DOMAIN IS THE 1004 00:38:30,720 --> 00:38:31,587 C-TERMINAL TRANSMEMBRANE DOMAIN 1005 00:38:31,587 --> 00:38:33,323 AND YOU END UP WITH INSTEAD OF 1 1006 00:38:33,323 --> 00:38:35,725 OR 2 LARGE HOLES, YOU EPPED UP 1007 00:38:35,725 --> 00:38:38,695 WITH ABOUT A THOUSAND PIN HOLES, 1008 00:38:38,695 --> 00:38:42,699 AND WE HAVE CRYOEM, AND CRYOEM, 1009 00:38:42,699 --> 00:38:45,835 NEGATIVE STAIN EM AND ALSO 1010 00:38:45,835 --> 00:38:47,270 ELECTRODYNAMICS MODELING AND ALL 1011 00:38:47,270 --> 00:38:48,171 SORTS OF BIOCHEMISTRY TO DEMON 1012 00:38:48,171 --> 00:38:49,072 TRAIT THIS IS RIGHT. 1013 00:38:49,072 --> 00:38:52,408 SO YOU END UP WITH A HEPTAMEROF 1014 00:38:52,408 --> 00:38:53,476 THE S105 PROTEIN AND ABOUT A 1015 00:38:53,476 --> 00:38:55,111 THOUSAND OF THEM EMPLOY NOW THE 1016 00:38:55,111 --> 00:38:56,913 PROBLEM IS, THAT'S TOO SMALL TO 1017 00:38:56,913 --> 00:38:58,047 LET THE LYSOSIEM OUT BUT THAT'S 1018 00:38:58,047 --> 00:39:00,350 NOT A PROBLEM HERE BECAUSE WHAT 1019 00:39:00,350 --> 00:39:02,785 HAPPENS IS, WHEN THESE FORM 1020 00:39:02,785 --> 00:39:04,320 THESE HOLES, THEY DEPOLARIZE THE 1021 00:39:04,320 --> 00:39:05,288 MEMBRANE CAN THAT ALLOWS THIS 1022 00:39:05,288 --> 00:39:06,723 MEMBRANE TO COME OUT OF THE 1023 00:39:06,723 --> 00:39:08,124 MEMBRANE WHICH ACTIVATES THE 1024 00:39:08,124 --> 00:39:11,694 ENZYME SO IT'S STILL A RAFT 1025 00:39:11,694 --> 00:39:13,162 CRITICAL CONCENTRATION DEPENDENT 1026 00:39:13,162 --> 00:39:19,001 STORY, JUST A DIFFERENT CHORIO 1027 00:39:19,001 --> 00:39:19,268 GRAPH. 1028 00:39:19,268 --> 00:39:21,771 SO THAT'S SHOWN BY THIS 1029 00:39:21,771 --> 00:39:22,572 PARALLEL. 1030 00:39:22,572 --> 00:39:23,673 HERE IS CANNONICLE SYSTEM WE'RE 1031 00:39:23,673 --> 00:39:28,111 TALKING ABOUT, AND HERE'S THE 1032 00:39:28,111 --> 00:39:29,045 NEW PINHOLE SYSTEM, THE ONLY 1033 00:39:29,045 --> 00:39:29,979 THING THAT CHANGE SYSTEM THE 1034 00:39:29,979 --> 00:39:31,514 SIZE OF THE HOLE AND THE RAFTS, 1035 00:39:31,514 --> 00:39:32,982 EVERYTHING ELSE IS THE SAME AND 1036 00:39:32,982 --> 00:39:34,617 THE GENETICS OF S21 ARE EQUALLY 1037 00:39:34,617 --> 00:39:36,119 CONVINCING AS THEY ARE FOR 1038 00:39:36,119 --> 00:39:36,352 LAMDA. 1039 00:39:36,352 --> 00:39:42,425 OKAY AND WE SHOULD HAVE NOTICED 1040 00:39:42,425 --> 00:39:43,793 THIS EARLY ON BECAUSE WHAT YOU 1041 00:39:43,793 --> 00:39:45,795 EXPECT SINCE IN THIS CASE, THE 1042 00:39:45,795 --> 00:39:47,029 ENDOLIES IS NOT COMING OUT 1043 00:39:47,029 --> 00:39:49,399 THROUGH 1 BIG HOLE, IT'S ALL THE 1044 00:39:49,399 --> 00:39:50,833 WAY AROUND THE HOLE ALREADY, SO 1045 00:39:50,833 --> 00:39:51,701 YOU'RE LOOKINGA THE DESTRUCTION 1046 00:39:51,701 --> 00:39:57,173 OF THE CELL WALL AND THAT'S WHAT 1047 00:39:57,173 --> 00:39:57,540 YOU SEE. 1048 00:39:57,540 --> 00:39:59,075 SO I WILL TAKE A RISK AND TRY TO 1049 00:39:59,075 --> 00:39:59,942 RUN THIS FORWARD MUCH THE 1050 00:39:59,942 --> 00:40:01,978 CONTROLLER OF BANKED 1S WILL ALL 1051 00:40:01,978 --> 00:40:08,217 GO ARK ROUND BEFORE THEY BLOW 1052 00:40:08,217 --> 00:40:11,287 UP. 1053 00:40:11,287 --> 00:40:13,156 AND NOTICE ALSO THAT ONLY 7 OF 1054 00:40:13,156 --> 00:40:15,425 10 CELLS THAT WE'RE HEARING HERE 1055 00:40:15,425 --> 00:40:17,059 DID THIS AND THIS TELLS US 1056 00:40:17,059 --> 00:40:17,994 SOMETHING WE ALREADY NOTICED 1057 00:40:17,994 --> 00:40:19,495 THAT THE PIN HOLE ASSESSMENTS 1058 00:40:19,495 --> 00:40:24,867 ARE NOT QUITE AS WELL TIMED AS 1059 00:40:24,867 --> 00:40:25,701 THE CANNONICLE WHOLE SYSTEMS 1060 00:40:25,701 --> 00:40:28,070 ARE, SO IF YOU DID THE LAMDA 1061 00:40:28,070 --> 00:40:29,872 S-PROTEIN WITH THIS, THEY WOULD 1062 00:40:29,872 --> 00:40:30,973 ALL BE LIESING WITHIN A 1063 00:40:30,973 --> 00:40:32,809 MACHINEUTE OR 2 O IT'S NOT QUITE 1064 00:40:32,809 --> 00:40:33,376 AS WELL CONTROLLED. 1065 00:40:33,376 --> 00:40:35,278 SO IN THE LAST 5 MINUTES HERE WE 1066 00:40:35,278 --> 00:40:38,414 WILL TALK ABOUT SPANINS, 2 OF 1067 00:40:38,414 --> 00:40:48,191 THE SPANIN MIEOF THERS ARE HERE, 1068 00:40:48,191 --> 00:40:53,763 IS IS HE THERE IN MANOY, ARE YOU 1069 00:40:53,763 --> 00:40:56,632 THERE IN OH WELL, SHE'S IN 1070 00:40:56,632 --> 00:40:57,900 [INDISCERNIBLE] LAB ANYWAY SO 1071 00:40:57,900 --> 00:40:59,535 MAYBE HE FELT LIKE HE KNEW IT 1072 00:40:59,535 --> 00:40:59,869 ALL. 1073 00:40:59,869 --> 00:41:01,637 BUT THESE ARE THE 4 PEOPLE WHO 1074 00:41:01,637 --> 00:41:04,273 DID THIS WORK, LIZ SUMMER DID 1075 00:41:04,273 --> 00:41:05,107 THE BIO-INFORMATICS AND 1076 00:41:05,107 --> 00:41:06,909 EVERYBODY ELSE DID GENETIC ANDS 1077 00:41:06,909 --> 00:41:07,243 BIOCHEMISTRY. 1078 00:41:07,243 --> 00:41:10,112 THIS IS THE WEIRDEST YEEN IN THE 1079 00:41:10,112 --> 00:41:11,280 WORLD WITH THE WEIRDEST PROTEINS 1080 00:41:11,280 --> 00:41:13,850 IN THE WORLD SO THE BOTTOM LINE 1081 00:41:13,850 --> 00:41:16,853 IS THE MAJOR PROTEIN, RZ PROTEIN 1082 00:41:16,853 --> 00:41:17,720 HAS A TRANSMEMORY RESPONSE BREAN 1083 00:41:17,720 --> 00:41:19,789 DOMAIN AND ABOUT A HUNDRED AMINO 1084 00:41:19,789 --> 00:41:22,458 OOH SIDS OF ALMOST TOTALLY ALPHA 1085 00:41:22,458 --> 00:41:24,060 HELICAL PROTEIN AND THEN 1086 00:41:24,060 --> 00:41:24,894 EMBEDDED IN THE DIFFERENT 1087 00:41:24,894 --> 00:41:28,097 READING FRAME IS THE RZ1 PROTEIN 1088 00:41:28,097 --> 00:41:29,832 WHICH ONLY HAS 40 RESIDUES IN 1089 00:41:29,832 --> 00:41:31,267 THE MATURE FORM POPULATED BY 1090 00:41:31,267 --> 00:41:34,237 PROLANES AND HAS AN OUTER 1091 00:41:34,237 --> 00:41:37,073 MEMBRANE LIPOPROTEIN SIGNAL 1092 00:41:37,073 --> 00:41:38,975 WHICH MEANS FOR THOSE WHO ARE 1093 00:41:38,975 --> 00:41:40,042 NOT FAMILIAR WITH LIPOPROTEINS, 1094 00:41:40,042 --> 00:41:42,144 BY THE WAY, BEFORE I GET TO--IT 1095 00:41:42,144 --> 00:41:48,784 TURNS OUT THAT PHAGE T1 HAS THE 1096 00:41:48,784 --> 00:41:50,019 SAME SYSTEM, PINHOLIN, AND 1097 00:41:50,019 --> 00:41:51,988 DIFFERENT TYPE OF SPANIN, THIS 1098 00:41:51,988 --> 00:41:53,489 CASE IT WAS [INDISCERNIBLE] BABY 1099 00:41:53,489 --> 00:41:55,324 WHILE HE WAS IN THE LABORATORY, 1100 00:41:55,324 --> 00:41:57,260 IT DOES HAVE A LIFE PROMOTING 1101 00:41:57,260 --> 00:42:00,329 SIGNAL BUT IT'S ALL BETA SHEET 1102 00:42:00,329 --> 00:42:05,568 PREDICTION AND HAS A C-TERMINAL 1103 00:42:05,568 --> 00:42:05,735 TMD. 1104 00:42:05,735 --> 00:42:07,003 SO THE BOTTOM LINE IS IN 1 CASE 1105 00:42:07,003 --> 00:42:11,140 YOU HAVE A PROTEIN COMPLEX THAT 1106 00:42:11,140 --> 00:42:13,009 SPANS THE ENTIRE PERIPLASM, IN 1107 00:42:13,009 --> 00:42:14,410 THIS CASE IT'S 2 PROTEINS ISSUES 1108 00:42:14,410 --> 00:42:18,114 THE 1 PROTEIN AND THE RZ1 1109 00:42:18,114 --> 00:42:19,682 PROTEIN, IN THE CASE OF T1 YOU 1110 00:42:19,682 --> 00:42:22,652 HAVE A SINGLE MOLECULE WITH A 1111 00:42:22,652 --> 00:42:23,252 COMPLETELY DIFFERENT TOPOLOGY 1112 00:42:23,252 --> 00:42:24,687 AND IT'S ALL IN THE SHEET, SO 1113 00:42:24,687 --> 00:42:26,622 NOTICE IN BOTH CASES THEY'RE 1114 00:42:26,622 --> 00:42:30,693 THREADED THROUGH THE GLYCAN MESH 1115 00:42:30,693 --> 00:42:34,497 WORK AND THEREFORE KOOBT DIFFUSE 1116 00:42:34,497 --> 00:42:34,764 LATERALLY. 1117 00:42:34,764 --> 00:42:36,265 AND THAT WE THINK IS HOW THEY'RE 1118 00:42:36,265 --> 00:42:36,566 CONTROLLED. 1119 00:42:36,566 --> 00:42:38,601 SO THE BOTTOM LINE IS, FIRST OF 1120 00:42:38,601 --> 00:42:39,936 ALL, WE IT SHOW THAT THREAR 1121 00:42:39,936 --> 00:42:50,513 REQUIRED AND SO I WILL START TE 1122 00:43:01,190 --> 00:43:01,357 CLOCK. 1123 00:43:01,357 --> 00:43:11,867 CAN I MAKE IT GO FASTER, THERE 1124 00:43:16,906 --> 00:43:17,306 WE GO. 1125 00:43:17,306 --> 00:43:20,876 YOU CAN GET THE WHOLE FORM, THE 1126 00:43:20,876 --> 00:43:22,411 OUTER MEMBRANE LEFT INTACT SO IT 1127 00:43:22,411 --> 00:43:24,513 TURNS INTO A FEAR AND WE ARE A 1128 00:43:24,513 --> 00:43:25,548 PICTURE OF THAT. 1129 00:43:25,548 --> 00:43:27,783 THIS THE ASSUME THING, FOR THE 1130 00:43:27,783 --> 00:43:30,219 SAME STORY, SO THIS IS A 1131 00:43:30,219 --> 00:43:32,288 CRYOPICTURE OF 1 OF THOSE CELLS 1132 00:43:32,288 --> 00:43:35,024 THAT HAS NO PANIN, THE OUTER 1133 00:43:35,024 --> 00:43:35,992 MEMBRANE IS INTACT AND HERE THE 1134 00:43:35,992 --> 00:43:37,893 NICE THING IS YOU CAN SEE, 1135 00:43:37,893 --> 00:43:40,062 HERE'S THE INNER MEMBRANE AND 1136 00:43:40,062 --> 00:43:41,364 THE ACTUAL S-HOLE, SO WE CAUGHT 1137 00:43:41,364 --> 00:43:42,898 IT IN THE ACT, 1 OF THE THINS 1138 00:43:42,898 --> 00:43:45,901 YOU LEARN FROM THIS IS THAT 1139 00:43:45,901 --> 00:43:48,037 ABSENT PHYSICAL STRESSES, THE 1140 00:43:48,037 --> 00:43:50,172 OUTER MEMBRANE IS CAPABLE IF 1141 00:43:50,172 --> 00:43:52,575 NOT--IF LEFT ALONE IS CAPABLE 1142 00:43:52,575 --> 00:43:54,777 FOR STANDING THE ENTIRE LINE OF 1143 00:43:54,777 --> 00:44:05,187 THE CELL WITHOUT--EACH WITHOUT 1144 00:44:05,187 --> 00:44:05,721 THE TBLI KAN--KANA. 1145 00:44:05,721 --> 00:44:07,056 OKAY O THIS IS WHAT WE THINK 1146 00:44:07,056 --> 00:44:12,962 WHAT HAPPENS WITH THE SINGLE OR 1147 00:44:12,962 --> 00:44:15,197 DOUBLE, ONCE THE ENDOLIESIN 1148 00:44:15,197 --> 00:44:18,601 DEGRADES THE FLI COLSIS CON, 1149 00:44:18,601 --> 00:44:19,035 WITH REARAINCHLMENT 1150 00:44:19,035 --> 00:44:19,969 OLIGMERRIZATION OCCURS AND THAT 1151 00:44:19,969 --> 00:44:22,038 LEADS TO COLLAPSE OF THE 1152 00:44:22,038 --> 00:44:24,540 MEMBRANE AND THESE 2 MODELS, WE 1153 00:44:24,540 --> 00:44:25,741 DON'T HAVE ANY EVIDENCE FOR 1154 00:44:25,741 --> 00:44:27,476 THESE, FOR THE ACTUAL FORMATION 1155 00:44:27,476 --> 00:44:30,346 OF THESE MODELS BUT WE DO HAVE 1156 00:44:30,346 --> 00:44:32,715 EVIDENCE FOR THE CLAPS IN 1157 00:44:32,715 --> 00:44:34,517 CONDENSATION OF THESE DOMAINS, 1158 00:44:34,517 --> 00:44:36,252 AND IF THIS LOOKS LIKE VIRAL 1159 00:44:36,252 --> 00:44:37,520 MEMBRANE PROEN TOOS TO YOU IF 1160 00:44:37,520 --> 00:44:38,821 YOU'RE INTO HA KIND OF THING, 1161 00:44:38,821 --> 00:44:41,891 THEY LOOK LIKE THE SAME TO US. 1162 00:44:41,891 --> 00:44:45,294 SO 1 STRONG POSITION OF 1163 00:44:45,294 --> 00:44:47,263 PREDICTION, IF YOU--IF YOU 1164 00:44:47,263 --> 00:44:48,230 EXTRAPOLATE THIS INTO 3 1165 00:44:48,230 --> 00:44:50,933 DIMENSIONS, WHAT THIS SAYS IS 1166 00:44:50,933 --> 00:44:53,869 THAT DURING LYSIS, THE LAST STEP 1167 00:44:53,869 --> 00:44:56,372 MODIFIED BY--MEDIATED BY SPANIN, 1168 00:44:56,372 --> 00:45:00,242 YOU OUGHT TO SEGGREGATE THE 1169 00:45:00,242 --> 00:45:01,477 PERIPLAZ MIDSIC CONTENTS IN 1170 00:45:01,477 --> 00:45:03,612 SECULAR FORM AND AT LEAST 1171 00:45:03,612 --> 00:45:04,914 INITIALLY THE PERIPLASMIC 1172 00:45:04,914 --> 00:45:06,115 CONTENTS DO NOT PARTICIPATE, 1173 00:45:06,115 --> 00:45:07,316 THEY'RE NOT RELEASED SO THAT'S 1 1174 00:45:07,316 --> 00:45:13,489 WAY WE CAN TEST THIS MODEL. 1175 00:45:13,489 --> 00:45:15,991 FIRST THIS IS TO SHOW FUSION 1176 00:45:15,991 --> 00:45:20,496 OCCURS, SO THIS IS--THIS IS 1177 00:45:20,496 --> 00:45:22,665 MAMOY, AND WHO ELSE DID THIS 1178 00:45:22,665 --> 00:45:24,200 WITH YOU, HE'S NOT HERE RIGHT 1179 00:45:24,200 --> 00:45:24,433 NOW. 1180 00:45:24,433 --> 00:45:26,102 SO THE BOTTOM LINE IS YOU CAN 1181 00:45:26,102 --> 00:45:27,303 IMET WITH THOSE--WE FOUND A 1182 00:45:27,303 --> 00:45:31,173 TRICK FOR PUTTING THE SPANIN 1183 00:45:31,173 --> 00:45:33,375 PERIPLASMIC DOMAINS ON THE 1184 00:45:33,375 --> 00:45:35,344 MEMBRANES, IT ALLOWS THEM BY 1185 00:45:35,344 --> 00:45:36,412 PUTTING DIFFERENT COLORS IN 1186 00:45:36,412 --> 00:45:37,480 DIFFERENT VESICLES WE CAN SEE 1187 00:45:37,480 --> 00:45:39,315 DIRECTLY WHETHER OR NOT THOSE 1188 00:45:39,315 --> 00:45:40,716 DOMAINS ARE CAPABLE OF FUSION IN 1189 00:45:40,716 --> 00:45:42,017 THE MEMBRANE AND IF THEY ARE, 1190 00:45:42,017 --> 00:45:45,688 AND IF YOU HAVE EITHER DEFECTIVE 1191 00:45:45,688 --> 00:45:50,760 MUTANTS OR SPANINS, YOU CAN 1192 00:45:50,760 --> 00:45:56,966 STABLE ABSORPTION BUT NO FUSION. 1193 00:45:56,966 --> 00:46:00,269 OKAY, SO LYSIS JEERN ROSTER, 2 1194 00:46:00,269 --> 00:46:04,240 TYPES OF HOLINs, 2 TYPES OF 1195 00:46:04,240 --> 00:46:05,207 ENDOLIESIN, THE CANNONICLE 1196 00:46:05,207 --> 00:46:06,342 ENDOLIESIN THERE ARE 2 TYPES OF 1197 00:46:06,342 --> 00:46:08,410 THOSE AND 2 TYPES OF SPANINS. 1198 00:46:08,410 --> 00:46:09,912 I DON'T KNOW WHY THERE'S ALL 1199 00:46:09,912 --> 00:46:11,147 THESE DIFFERENT POSSIBILITIES 1200 00:46:11,147 --> 00:46:13,682 AND WE'VE EITHER FOUND IN NATURE 1201 00:46:13,682 --> 00:46:16,152 OR FOUND OR CONSTRUCTED IN A 1202 00:46:16,152 --> 00:46:18,053 LABORATORY ALMOST EVERY 1203 00:46:18,053 --> 00:46:19,355 COMBINATION OF THESE 3 1204 00:46:19,355 --> 00:46:21,190 COMPONENTS BUT THAT'S NOT ALL. 1205 00:46:21,190 --> 00:46:25,694 AND WE PROMISE TO JUF MENTION 1206 00:46:25,694 --> 00:46:26,829 ANTIHOLIN, ABOUT HALF THE 1207 00:46:26,829 --> 00:46:28,497 SYSTEMS WE STUDY HAVE 1208 00:46:28,497 --> 00:46:29,365 ANTIHOLINs I DON'T HAVE TIME 1209 00:46:29,365 --> 00:46:32,101 TO TALK ABOUT WHAT THEY DO BUT 1210 00:46:32,101 --> 00:46:33,903 IN EACH CASE, 1 ACTIVATES 1 1211 00:46:33,903 --> 00:46:35,404 HOLIN FOR THE PURPOSE OF THE 1212 00:46:35,404 --> 00:46:36,672 LYSIS CLOCK AND THE MOST 1213 00:46:36,672 --> 00:46:39,642 INTERESTING 1 IS T4 WHICH HAS A 1214 00:46:39,642 --> 00:46:42,411 AS YOU REMEMBER IT HAS A 1215 00:46:42,411 --> 00:46:44,647 CYTOPLASMIC DOMAIN AND A 1216 00:46:44,647 --> 00:46:45,114 PERIPLASMIC DOMAIN. 1217 00:46:45,114 --> 00:46:48,083 IT TURPS OUT THERE ARE 2 1218 00:46:48,083 --> 00:46:53,789 DIFFERENT HOLEINS A SOLUBLE 1 1219 00:46:53,789 --> 00:46:57,026 AND AND OACH MORE REMARKABLY, 1220 00:46:57,026 --> 00:46:57,827 THE CONTEXT BETWEEN THOSE AND 1221 00:46:57,827 --> 00:47:00,696 THE HOLEIN AND THE T-4 CASE THAT 1222 00:47:00,696 --> 00:47:03,065 COMPLEX IS DENSATIVE TO DNA IN 1223 00:47:03,065 --> 00:47:09,338 THE PERIPLASMA, SO THE MODEL IS 1224 00:47:09,338 --> 00:47:10,773 THAT IF YOU INRECYCLING, IF YOU 1225 00:47:10,773 --> 00:47:12,708 HAVE A SECONDARY PHAGE INFECTING 1226 00:47:12,708 --> 00:47:14,376 DNA, THE DNA GOES IN, IF YOU 1227 00:47:14,376 --> 00:47:16,478 PERMIT THAT DMA FROM GETTING TO 1228 00:47:16,478 --> 00:47:18,981 THE CYTOPLASM, IT THEN CAN BE 1229 00:47:18,981 --> 00:47:22,651 ACT AS A HARBINGER THE 1230 00:47:22,651 --> 00:47:24,019 PARTICULAR INFECKED T4 CELL IT'S 1231 00:47:24,019 --> 00:47:26,021 IN AN REQUIREMENT WHERE IT'S NOT 1232 00:47:26,021 --> 00:47:28,924 A GOOD IDEA TO LYSE. 1233 00:47:28,924 --> 00:47:29,992 BECAUSE IT'S BEING SUPER 1234 00:47:29,992 --> 00:47:33,629 INFEBBED SO A DNA BINDS TO IT, 1235 00:47:33,629 --> 00:47:35,264 STABILIZES AND PREVENTS THE 1236 00:47:35,264 --> 00:47:37,466 HOLIN FROM AG TBREIGATING INTO 1237 00:47:37,466 --> 00:47:39,535 HOLES AND TRIGGERING LYSIS, SO 1238 00:47:39,535 --> 00:47:41,537 IT'S AN INTERESTING STORY IN ITS 1239 00:47:41,537 --> 00:47:43,772 OWN WRITE AND GOOD 1240 00:47:43,772 --> 00:47:45,908 CRYSTALLOGRAPHY BY WHY 1241 00:47:45,908 --> 00:47:46,742 IMPEDIMENTS' LAB AND IN 1242 00:47:46,742 --> 00:47:47,910 PARTICULAR FOR THAT UP. 1243 00:47:47,910 --> 00:47:50,779 SO THIS I WANT TO SHOW YOU WHAT 1244 00:47:50,779 --> 00:47:51,714 WE JUST PRELIMINARY EXPERIMENTS 1245 00:47:51,714 --> 00:47:55,017 WE TRY TO SHOW WHETHER OR NOT 1246 00:47:55,017 --> 00:47:58,287 IT'S TRUE, THAT AFTER HISPANIN 1247 00:47:58,287 --> 00:48:00,623 FUNCTION THE PERIPLASMIC 1248 00:48:00,623 --> 00:48:01,557 CONTENTS ARE SEGREGATED. 1249 00:48:01,557 --> 00:48:03,125 SO IN THIS EXPERIMENT WE 1250 00:48:03,125 --> 00:48:08,597 SECRETED THE SUPER FOLDER GFP TO 1251 00:48:08,597 --> 00:48:10,032 THE PERIPLASM, USING A SIGNAL 1252 00:48:10,032 --> 00:48:11,433 SEQUENCE AND THEN LET IT TAKES 1253 00:48:11,433 --> 00:48:11,767 IT COURSE. 1254 00:48:11,767 --> 00:48:13,402 THIS IS A REGULAR LAMDA LYSIS 1255 00:48:13,402 --> 00:48:22,745 WITH ALL THE GENES NOW WE'RE 1256 00:48:22,745 --> 00:48:25,114 WATCHING GFP IN THE PERIPLASM, 1257 00:48:25,114 --> 00:48:27,349 AND IT SHOULD STAY IN THE 1258 00:48:27,349 --> 00:48:28,984 PERIPLASM, BECAUSE IT'S NOT 1259 00:48:28,984 --> 00:48:39,495 ATTACHED TO ANY LYSIS PROTEIN. 1260 00:48:44,600 --> 00:48:45,701 SO--SO WHAT'S HAPPENING IS WE 1261 00:48:45,701 --> 00:48:47,503 THINK VERY NEAR THE END OF LYSIS 1262 00:48:47,503 --> 00:48:51,173 AS THE PEPTIDE O O GLYCAN IS 1263 00:48:51,173 --> 00:48:54,310 DESTROYED, THE PERIPLASM IS 1264 00:48:54,310 --> 00:48:55,411 SQUISHED OUT INTO THE HOLES OF 1265 00:48:55,411 --> 00:48:56,845 THE CELL, I DON'T KNOW THE 1266 00:48:56,845 --> 00:48:58,047 PHYSICS OF IT, BUT AS YOU CAN 1267 00:48:58,047 --> 00:48:59,915 SEE, AT LEAST IN THIS--BOTH 1268 00:48:59,915 --> 00:49:02,384 CASES, LET ME BACK THIS UP, 1 OF 1269 00:49:02,384 --> 00:49:03,852 THEM KIND OF FELL APART AFTER A 1270 00:49:03,852 --> 00:49:05,654 WHILE, BUT ALL OF THE INTENSITY 1271 00:49:05,654 --> 00:49:07,556 THAT WAS THERE, IN THE 1272 00:49:07,556 --> 00:49:09,258 PERIPHERY, IS NOW THERE THIS 1273 00:49:09,258 --> 00:49:12,528 THOSE 2 VESICLES AND JUST TO 1274 00:49:12,528 --> 00:49:14,630 MAKE SURE THAT THIS ISN'T AN 1275 00:49:14,630 --> 00:49:17,666 ARTIFACT, WE WENT ON TO SHOW 1276 00:49:17,666 --> 00:49:20,035 THAT IF YOU GET RID OF THE 1277 00:49:20,035 --> 00:49:21,103 SPANINS, NOW THERE'S NEVER GOING 1278 00:49:21,103 --> 00:49:23,772 TO BE A FUSION OF THE INNER OR 1279 00:49:23,772 --> 00:49:24,606 OUTER MEMBRANE. 1280 00:49:24,606 --> 00:49:27,910 JUST HAVE A HOLE IN THE 1281 00:49:27,910 --> 00:49:29,078 ENDOLIESIN FUNCTION, SAME 1282 00:49:29,078 --> 00:49:30,045 EXPERIMENTAL DETAIL, SO NOW OF 1283 00:49:30,045 --> 00:49:31,547 COURSE YOU END UP WITH THE 1284 00:49:31,547 --> 00:49:32,748 SPHEREICAL CELL AND AGAIN 1285 00:49:32,748 --> 00:49:33,816 WITHOUT THE PEPTIDE O O FLI 1286 00:49:33,816 --> 00:49:35,651 COLSIS KAN--KANA, FOR SOME 1287 00:49:35,651 --> 00:49:37,219 REASON THE PERIPLASM IS ALL 1288 00:49:37,219 --> 00:49:39,021 SQUISHED INTO 1 DOMAIN ISSUE THE 1289 00:49:39,021 --> 00:49:40,322 1 PART WE'RE NOT QUITE 1290 00:49:40,322 --> 00:49:42,858 UNDERSTANDING IS WHY IT'S 1291 00:49:42,858 --> 00:49:44,026 LEAKING GFP BACK INTO THE 1292 00:49:44,026 --> 00:49:46,428 CYTOPLASM AND THERE MIGHT BE 1293 00:49:46,428 --> 00:49:47,930 SMALL S-HOLES MOST OF THE HOLES 1294 00:49:47,930 --> 00:49:49,665 OF THE HOLEIN IS IN LARGE HOLES 1295 00:49:49,665 --> 00:49:52,001 AND MAYBE A FEW OF SMALL HOLES 1296 00:49:52,001 --> 00:49:55,738 SO IT ALLOWS GFP TO LEAK BACK IN 1297 00:49:55,738 --> 00:49:56,705 BUT WE HAVEN'T DONE ENOUGH 1298 00:49:56,705 --> 00:49:59,608 EXPERIMENTS TO DEAL WITH THAT 1299 00:49:59,608 --> 00:49:59,908 YET. 1300 00:49:59,908 --> 00:50:02,211 OKAY, SO THE LAST SLIDE IS WHAT 1301 00:50:02,211 --> 00:50:05,748 I KAW LYSIS EFTHIMIOS MOLOGY, 1302 00:50:05,748 --> 00:50:11,920 WHY DO HOLINs EXIST AT ALL 1303 00:50:11,920 --> 00:50:13,489 EXCEPT IT GIVE ME 40 YEARS OF 1304 00:50:13,489 --> 00:50:15,224 FUNDING BUT THAT'S GOOD ENOUGH 1305 00:50:15,224 --> 00:50:15,824 FOR ME. 1306 00:50:15,824 --> 00:50:17,559 BUT PUT A SIGNAL AND SECRETE IT 1307 00:50:17,559 --> 00:50:21,463 WILL DO WHATEVER YOU NEED TO DO 1308 00:50:21,463 --> 00:50:24,299 AND SO, THAT'S POSSIBLE AND IT 1309 00:50:24,299 --> 00:50:30,973 TURNS OUT IN 1 OF THE MOST 1310 00:50:30,973 --> 00:50:32,508 COLOSSAL HYPOTHESIS CANCELING 1311 00:50:32,508 --> 00:50:35,778 THAT I'VE EVER--I LOOKED AT THE 1312 00:50:35,778 --> 00:50:37,112 SEQUENCE OF THE MUSE GENOME, 1 1313 00:50:37,112 --> 00:50:38,414 OF THE CLASSIVE PHAGES, 1 OF THE 1314 00:50:38,414 --> 00:50:39,848 LAST 1S TO HAVE THE LOISIS 1315 00:50:39,848 --> 00:50:43,352 STUDIED AND THERE WAS A CLEAR 1316 00:50:43,352 --> 00:50:44,186 HOLEIN AND ANTIHOLEIN, RIGHT 1317 00:50:44,186 --> 00:50:46,455 NEXT TO THE LYSIS YEENS AND SO I 1318 00:50:46,455 --> 00:50:50,626 ACTUALLY LABELED IN THE REVOW, 1319 00:50:50,626 --> 00:50:52,628 THIS LIKELY HOLEIN AND ANTIHOLIM 1320 00:50:52,628 --> 00:50:54,396 AND IT TURNS OUT THEY'RE NOT 1321 00:50:54,396 --> 00:50:57,232 RELATED TO LYSIS AND IT HAS A 1322 00:50:57,232 --> 00:50:58,434 ENDOLYSIS AND HAS A SPANNED 1323 00:50:58,434 --> 00:51:00,903 SYSTEM BUT IT DOESN'T HAVE A 1324 00:51:00,903 --> 00:51:05,074 HOLIN, SO HAD THIS CASE, THE USE 1325 00:51:05,074 --> 00:51:08,177 SYSTEM USES AN ANTIENDOLIESIN 1326 00:51:08,177 --> 00:51:10,679 THAT PREVENTS THE ACTIVITY OF 1327 00:51:10,679 --> 00:51:11,947 THE ENDOLIESIN AFTER SECRETION. 1328 00:51:11,947 --> 00:51:15,551 SO THIS IS WORK THAT'S BEING 1329 00:51:15,551 --> 00:51:17,319 DONE BY JO LEAN RAMSEY WHO JUST 1330 00:51:17,319 --> 00:51:19,188 SET HER LAB UP AT TEXAS A&M AND 1331 00:51:19,188 --> 00:51:22,724 CARRYING ON A LOT OF MY 1332 00:51:22,724 --> 00:51:26,595 MULTIYEEN LICENSE WORK. 1333 00:51:26,595 --> 00:51:33,102 AND EVEN MORE SIMPLE IS WHY NOT 1334 00:51:33,102 --> 00:51:36,405 USE THE SINGLE GENE LYSIS 1335 00:51:36,405 --> 00:51:37,106 AUTOLYSIS SYSTEM? 1336 00:51:37,106 --> 00:51:39,508 SO WE'VE BEEN TALKING ABOUT THIS 1337 00:51:39,508 --> 00:51:43,712 SYSTEM, WE CALL IT MULTIGENE 1338 00:51:43,712 --> 00:51:47,149 LYSIS, BUT ALL SMALL PHAGES LIKE 1339 00:51:47,149 --> 00:51:50,219 ALL SINGLE STRAND PHAGES AND 1340 00:51:50,219 --> 00:51:52,187 SINGLE STRAND RNAA PHAGES, THEY 1341 00:51:52,187 --> 00:51:53,722 ACCOMPLISH THIS WITH YEEPS, 1342 00:51:53,722 --> 00:51:55,991 THESE YEENS ARE INHIBITING STEPS 1343 00:51:55,991 --> 00:51:57,392 IN THE CELL WALL SIJT SIS AND 1344 00:51:57,392 --> 00:51:59,261 STIEPS NOT. 1345 00:51:59,261 --> 00:52:01,230 SO, BUT WHATEVER THEY DO, 1346 00:52:01,230 --> 00:52:03,532 THERE'S NO ENZYME, CARRIED BY 1347 00:52:03,532 --> 00:52:09,605 THE BACTERIA PHAGE SO INSTEAD 1348 00:52:09,605 --> 00:52:10,806 THESE ARE AUTOLYSIS EVENTS WE 1349 00:52:10,806 --> 00:52:12,441 SHOW IF YOU IMK OUT THE LAMDA 1350 00:52:12,441 --> 00:52:16,845 AND REPLACE IT WITH THE E-YEEN 1351 00:52:16,845 --> 00:52:20,749 OF 5 FORWHICH WE KNOW WORKS BY 1352 00:52:20,749 --> 00:52:22,484 INHIBITING THE GLYCAN, LAMDA 1353 00:52:22,484 --> 00:52:23,552 MAKES PERFECTLY GOOD PLAQUES SO 1354 00:52:23,552 --> 00:52:25,187 AT LEAST AT THE LEVEL OF PLAQUE 1355 00:52:25,187 --> 00:52:28,824 FORM EGG THE LAMDA EGENE COMP 1356 00:52:28,824 --> 00:52:30,792 LEAPTS OF DEFECT, SO--BUT YET, 1357 00:52:30,792 --> 00:52:32,694 SO FAR AS FAR AS WE CAN TELL, 1358 00:52:32,694 --> 00:52:35,430 ALL DOUBLE STRAND DNA FAINLS USE 1359 00:52:35,430 --> 00:52:38,634 THE SYSTEM AND SUSPECT THEY MUST 1360 00:52:38,634 --> 00:52:39,301 BE POWERFUL EVOLUTIONARY FORCES 1361 00:52:39,301 --> 00:52:41,203 TO MAKE THAT HAPPEN. 1362 00:52:41,203 --> 00:52:42,404 AGAIN, WE'RE BACK TO THE PICTURE 1363 00:52:42,404 --> 00:52:44,606 OF THE PEOPLE WHO DID IT ALL AND 1364 00:52:44,606 --> 00:52:46,708 I APPRECIATE YOUR PATIENCE. 1365 00:52:46,708 --> 00:52:54,449 [ APPLAUSE ] 1366 00:52:54,449 --> 00:53:01,123 ANYONE HERE IS FREE TO USE THE 1367 00:53:01,123 --> 00:53:03,258 MIC'S FOR QUESTIONS AS WE WAIT 1368 00:53:03,258 --> 00:53:03,492 ONLINE. 1369 00:53:03,492 --> 00:53:06,862 NTHANK . 1370 00:53:06,862 --> 00:53:09,231 >> THANK YOU VERY MUCH. 1371 00:53:09,231 --> 00:53:09,765 FASCINATING. 1372 00:53:09,765 --> 00:53:15,103 MY MIND WENT ON WHEN THEY ARE 1373 00:53:15,103 --> 00:53:20,542 BURSTING IS THERE ANY, ANY 1374 00:53:20,542 --> 00:53:23,779 ENERGY DISPENSED AND/OR CAN YOU 1375 00:53:23,779 --> 00:53:27,015 PUT THESE UNDER HIPOXIC OR HYPER 1376 00:53:27,015 --> 00:53:28,850 OXIC CONDITION TO SEE WHETHER 1377 00:53:28,850 --> 00:53:32,621 THEY CHANGE THE TIMING OF THE 1378 00:53:32,621 --> 00:53:32,955 BURSTING? 1379 00:53:32,955 --> 00:53:34,156 >> THE ONLY THING THAT MATTER 1380 00:53:34,156 --> 00:53:35,123 SYSTEM THE MEMBRANE POTENTIAL 1381 00:53:35,123 --> 00:53:36,892 BUT THERE'S A LOT OF WAYS CAN 1382 00:53:36,892 --> 00:53:40,028 YOU MESS UP THE MEMBRANE. 1383 00:53:40,028 --> 00:53:41,797 FOR EXAMPLE, EVEN IN 1384 00:53:41,797 --> 00:53:43,966 [INDISCERNIBLE] LABORATORY BEAN 1385 00:53:43,966 --> 00:53:47,469 IN THE 1800S, WE KNEW FOR A LONG 1386 00:53:47,469 --> 00:53:50,505 TIME IF YOU INFECT E.COLI WITH 1387 00:53:50,505 --> 00:53:52,341 ANYTHING, ANY LAMDA T-4 ANY 1388 00:53:52,341 --> 00:53:54,109 PHAGE, YOU CAN'T TOP THE SHAKER, 1389 00:53:54,109 --> 00:53:55,410 YOU CAN'T SPIN THEM DOWN AND 1390 00:53:55,410 --> 00:53:57,212 THAT'S BECAUSE IF YOU STOP THE 1391 00:53:57,212 --> 00:53:59,748 SHAKER FLASK IN DENSE CULTURE 1392 00:53:59,748 --> 00:54:02,517 LIKE OD.2, THE ATTENTION DROPS 1393 00:54:02,517 --> 00:54:03,585 IMMEDIATELY BECAUSE IT'S BEING 1394 00:54:03,585 --> 00:54:05,053 SUCKED UP BY ALL THE CELLS, SO 1395 00:54:05,053 --> 00:54:06,255 THE BOTTOM LINE IS, ANYTHING 1396 00:54:06,255 --> 00:54:07,956 THAT ASHINGS THE MEMBRANE 1397 00:54:07,956 --> 00:54:10,158 POTENTIAL AND WE'VE SHOWN USING 1398 00:54:10,158 --> 00:54:11,426 THAT, SPINNING BUG EXPERIMENT 1399 00:54:11,426 --> 00:54:14,596 THAT WE SHOWED, THE YOU CAN USE 1400 00:54:14,596 --> 00:54:16,265 THAT TO MEASURE THE MEMBRANE 1401 00:54:16,265 --> 00:54:17,799 POTENTIAL AND AT WHICH TIME IT 1402 00:54:17,799 --> 00:54:19,434 TRIGGERS AND IT ONLY REQUIRES A 1403 00:54:19,434 --> 00:54:21,536 30% REDUCTION IN MEMBRANE 1404 00:54:21,536 --> 00:54:22,604 POTENTIAL FROM ABOUT 150 MILLI 1405 00:54:22,604 --> 00:54:24,039 VOWELS DOWN TO LIKE 100 AND 1406 00:54:24,039 --> 00:54:25,340 THAT'S ENOUGH TO TRIGGER. 1407 00:54:25,340 --> 00:54:31,313 SO YOU CAN GROW, CAN YOU DO 1408 00:54:31,313 --> 00:54:32,748 THESE EXPERIMENTS IN ANOXIC 1409 00:54:32,748 --> 00:54:36,652 CONDITIONS AS LONG AS YOU'RE 1410 00:54:36,652 --> 00:54:38,186 FERMENITATING THE METHODS. 1411 00:54:38,186 --> 00:54:48,463 NCOULD THEY BEHAVE SOMEWHAT LIKE 1412 00:54:48,463 --> 00:54:49,865 MITOCHONDRIAOXIDATIVE PROCESS. 1413 00:54:49,865 --> 00:54:54,970 >> I AGREE, BCL IS 1 OF THE 1414 00:54:54,970 --> 00:54:56,605 PROTEINS IN THE MITOGENIC 1415 00:54:56,605 --> 00:55:00,142 PATHWAY, IT TURNS OUT THE BCL 1416 00:55:00,142 --> 00:55:01,410 PROTEIN COMPLEMENTINGS LAMDA S, 1417 00:55:01,410 --> 00:55:04,579 SO MY NAME IS ON A PAPER THAT 1418 00:55:04,579 --> 00:55:06,214 SAYS THOSE PROTEINSOR HOLINs, 1419 00:55:06,214 --> 00:55:07,783 BUT IT'S NOT TRUE BECAUSE THEY 1420 00:55:07,783 --> 00:55:10,686 DON'T HAVE THE--THE KEY ELEMENT 1421 00:55:10,686 --> 00:55:13,522 OF THE HOLINs IS THEY HAVE TO 1422 00:55:13,522 --> 00:55:14,356 BE TRIGGABLE BY THE POISON 1423 00:55:14,356 --> 00:55:16,525 BECAUSE WE THINK THAT'S INTEGRAL 1424 00:55:16,525 --> 00:55:18,060 TO THE MECHANISM AND THE WHOLE 1425 00:55:18,060 --> 00:55:21,029 TIMING THING, NOTHING WILL WORK 1426 00:55:21,029 --> 00:55:23,398 WITHOUT THAT, BUT I COULDN'T 1427 00:55:23,398 --> 00:55:26,301 TURN DOWN GETTING A PAPER. 1428 00:55:26,301 --> 00:55:27,135 >> VERY NICE, THANK YOU. 1429 00:55:27,135 --> 00:55:33,175 >> I HAVE A QUESTION OVER HERE. 1430 00:55:33,175 --> 00:55:33,542 >> YEAH. 1431 00:55:33,542 --> 00:55:37,312 >> HI, HOW DO THE HOLINS GO TO 1432 00:55:37,312 --> 00:55:39,614 THE CURVEATURE OR TO THE POLES. 1433 00:55:39,614 --> 00:55:40,549 >> THEY DON'T ALWAYS DO THAT I 1434 00:55:40,549 --> 00:55:42,417 WAS SORT OF PISSED OFF THAT 80% 1435 00:55:42,417 --> 00:55:43,952 OF THEM HAPPEN AT THE POLES BUT 1436 00:55:43,952 --> 00:55:46,188 THEY DON'T ALWAYS DO THAT, I 1437 00:55:46,188 --> 00:55:48,123 THINK ITIA A DECISION EARLY ON 1438 00:55:48,123 --> 00:55:49,458 BECAUSE WHEN THEY SHOW UP 1439 00:55:49,458 --> 00:55:51,193 ELWHERE, THEY DON'T SHOW UP AT 1440 00:55:51,193 --> 00:55:52,594 THE POLES, EITHER 1 SO I SUSPECT 1441 00:55:52,594 --> 00:55:56,131 THERE'S SOME SORT OF SORTING 1442 00:55:56,131 --> 00:55:57,866 EVENT THAT OCCURS, FOR IT'S NOT 1443 00:55:57,866 --> 00:56:00,135 LYSIS SPECIFIC, THERE'S A LOT OF 1444 00:56:00,135 --> 00:56:02,037 STILL A LOT OF CONTROVERSY ABOUT 1445 00:56:02,037 --> 00:56:04,005 HOW ANY PROTEINS FINDS THE 1446 00:56:04,005 --> 00:56:04,206 POLES. 1447 00:56:04,206 --> 00:56:08,443 THE ANSWER IS I DON'T KNOW. 1448 00:56:08,443 --> 00:56:13,648 NWELL, THERE ARE NO QUESTIONS 1449 00:56:13,648 --> 00:56:14,082 ONLINE SURPRISINGLY. 1450 00:56:14,082 --> 00:56:18,053 LOTS OF PEOPLE WATCHING SO 1451 00:56:18,053 --> 00:56:18,720 DR. LORSCH--I'M SORRY, SOMETHING 1452 00:56:18,720 --> 00:56:20,288 CAME IN, THAT WAS NOTHING, THAT 1453 00:56:20,288 --> 00:56:22,891 WAS MY TEST TO SEE IF IT WAS 1454 00:56:22,891 --> 00:56:23,125 WORKING. 1455 00:56:23,125 --> 00:56:24,292 >> ACTUALLY SINCE THIS IS A 1456 00:56:24,292 --> 00:56:29,064 LECTURE THERE IS ACTUALLY A 1457 00:56:29,064 --> 00:56:31,099 TEST. 1458 00:56:31,099 --> 00:56:32,734 [LAUGHTER] 1459 00:56:32,734 --> 00:56:35,637 LET'S THANK RY FOR A 1460 00:56:35,637 --> 00:56:38,840 FANTASTIC TOUR DEFORCE TALK. 1461 00:56:38,840 --> 00:56:41,243 AND THERE WILL BE A SHORT 1462 00:56:41,243 --> 00:56:43,044 RECEPTION OUTSIDE FOR THOSE WHO 1463 00:56:43,044 --> 00:56:45,447 ARE HERE IN REAL LIFE AND THOSE 1464 00:56:45,447 --> 00:56:47,149 OF YOU WHO ARE VIRTUAL, YOUR ON 1465 00:56:47,149 --> 00:56:49,351 YOUR OWN, GO GET A COOKIE DOWN 1466 00:56:49,351 --> 00:56:50,519 STAIRS OR WHATEVER YOU WANT TO 1467 00:56:50,519 --> 00:56:55,624 DO BUT THANKS AGAIN RY. 1468 00:56:55,624 --> 00:57:05,834 [ APPLAUSE ]