Chaperone Newsletter featuring LIFE of PROTEINS no.14 CONTENTS Announcement Information Issue of the Issue Interview Inseider Story Essay Mini Review

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1 featuring 2005 No

2 Chaperone Newsletter featuring LIFE of PROTEINS no.14 CONTENTS Announcement Information Issue of the Issue Interview Inseider Story Essay Mini Review Meeting Report Replay Calendar Epilogue

3 Announcement March 2005 No.14 1

4 Information 4 q2w q1w ,600,000 q2w q3w 9 11 q4w q3w q4w Translation and folding Translocation and transport Quality control and degradation Disorder and diseases 4 Dr. Walter Neupert 3 dinner talk 7 invited speaker q5w 13 q1w q1w

5 q2w q3w q1w q2w q3w International Conference Life of Proteins CREST CREST 1 Translation and folding: 6 B. Bukau Universitat Heidelberg, Heidelberg U. F. Hartl Max-Planck-Institut fur Biochemie, Martinsried A. Horwich Yale University, New Haven R. Sauer Massachusetts Inst. Technol., Cambridge I. Braakman Utrecht University, Utrecht E. Craig University of Wisconsin 2 Translocation and transport: 6 A. Johnson Texas A&M University, College Station W. Neupert Ludwigs-Maximilians University, Munich N. Pfanner Universitaet Freiburg, Freiburg T. Rapoport Harvard Medical School, Boston March 2005 No.14 3

6 J. Soll Ludwig-Maximilians University, Munich R. Erdmann Ruhr-Universit, Bochum 3 Quality control and degradation: 6 R. Kopito Stanford University, Stanford P. Walter J. Weissman University of California-San Francisco, San Francisco T. Sommer Max-Delbruck-Center, Berlin D. Ron New York University Medical Center, New York D. Ng Pennsylvania State University, Pennsylvania 4 Disorder medical aspect): 5 R. Morimoto Northwestern University, Chicago R. C. Austin McMaster Univ., Hamilton, Canada W. Paschen Max-Planck Inst., Koeln) J. Yuan Harvard Medical School, Boston) E. Rugarli Telethon Institute of Genetics and Medicine, Italy D. Cyr University of North Carolina, Chapel Hill URL pdf

7 Issue of the Issue in vitro Issue of The Issue SecYEG/Sec Archaea SecYE Sec61 SecA SecG co-translational SecY Sec61 Y/61 Y/61 10 TM E/ G/ 1 3 TM 2 closed3.2 1 q1wy TM1-5 TM6-10 SecE q2w20 q3wy March 2005 No.14 5

8 TM7-TM2b TM2a Y Lateral gate SecY Y q4w 5 8 Pore ring Pore ring q5wtm2a Pore ring Pore q6w TM2 TM3 TM7 TM8 Hetero-trimer 2 4 Rapoport 2 Back to back 2 side by side SecA Y TM in vitro SecY

9 SecE E Y E SecA SecY SecE YE SecA 60 Johnson Johnson BiP Rapoport Johnson BiP Rapoport Partition White sequential point of view concerted scheme 3 4 Truncated mrna Spiess Lateral gate Dead-end Partition 10 March 2005 No.14 7

10 Spiess N N Topogenic signal Rapoport Topogenic signal Topogenic signal Pore Spiess Sec61 2 TM2a 1 TM8 Positive inside rule TM Rapoport Lateral gate TM Partition White Topogenic signal TM Lateral gate TRAM YidC Membrane chaperone SecY Lateral gate TM2b 7 SecA insertion SecA SecA insertion SecYEG SecA SecG SecA TRAM YidC Sec62/Sec63, TRAP, SecDF/YajC, OST Further reading: Clelmons, W.M.Jr., Rapoport, T.A.: Structural insight into the protein translocation channel. Curr. Opinion Struct. Biol. 14, Rapoport, T.A., et al.: Membrane-protein integration and the role of the translocation channel. Trends Cell Biol. 14, Alder, N.N. and Johnson, A.E.: Cotranslational membrane protein biogenesis at the endoplasmic reticulum. J. Biol. Chem. 279, White, S.H.: Translocon, thermodynamics, and the folding of membrane proteins. FEBS Lett. 555, White, S.H. and von Heijne, G.: The machinery of membrane protein assembly. Curr. Opinion Struct. Biol. 14, Higy, M., Junne, T. and Spiess, M.: Topogenesis of membrane proteins at the endoplasmic reticulum. Biochemistry 43, Biochim. Biophys. Acta 1694, no. 1: Protein Export/Secretion in Bacteria (2004) Pmf Pmf Pmf 8

11 Interview 3 90 Cell March 2005 No.14 9

12 Norman Davidson DNA TV LA 5 U

13 12 21 trna ESR trna Lys-tRNA trna Peter von Hippel 1969 trna John Menninger trna trna trna trna trna trna March 2005 No.14 11

14 mrna AUGAAU 5 82 JMB Charles Cantor UCSF PhD T 2 T Charlie trna f-met-trna Phe-tRNA L 4 f-met-trna P Phe-tRNA A trna 20 3A in vitro SRP trna EF-Tu trna 4 trna EF-Tu trna trna EF-Tu trna trna trna - trna EF-Tu 17 12

15 Peter Walter NMR Jeff 10 A&M Jeff Schatz Jeff Jeff 80 SRP Blobel trna Jeff Jeff Blobel PhD UCSF Peter Walter SRP trna 2 UCSF 4 SRP SRP 84 SRP54 10 NBD 50cm March 2005 No.14 13

16 A&M 1993 Cell Cell 73, Cell Alex Spirin 4 Cell Cell Cell 10 NIH Cell Northwestern Cell 1 Cell Cell 5 14

17 ER 3 Cell 116, FRET Insider Story 2004 March 2005 No.14 15

18 2004 The discovery of ubiquitinmediated protein degradation - Aaron Ciechanover Avram Hershko Irwin Rose 3 1 Winner Winner NHK 10 4 Hershko 16

19 Winners Hershko Ciechanover Alexander Varshavsky 3 Varshavsky Rose everywhere Winner Life of Proteins 12, ATP March 2005 No.14 17

20 Albert Lasker Award for Basic Medical Research The discovery of ubiquitin pathway Hershko Ciechanover Varshavsky Regulation of cellular functions by the ubiquitin-proteasome system UPS Nobel Forum Plenary Lecture Hershko Varshavsky Hershko Varshavsky Hershko 3 Ciechanover Hershko ATP Alfred Goldberg Rose Varshavsky Irwin Rose 1 2 Rose Avram Hershko Hershko 70 Lasker Wolf Hershko 18

21 sabbatical Hershko Plenary Lecture 1978 Hershko Hershko Aaron Ciechanover Hershko Ciechanover Ciechanover Hershko Ciechanover Irwin Rose ATP Mel Simpson 1953 ATP 1976 NIH Hershko Ciechanover Hershko Alfred L. Goldberg ATP 1977 PNAS 1977Hershko Rose sabbatical Hershko Ciechanover sabbatical Rose 1978 ATP Fraction I Fraction II ATP Ciechanover Fraction I APF-1 ATP-dependent proteolysis factor APF-1 APF-1 Rose ATP E1 pyrophosphate exchange, thiol ester, ubiquitin aldehyde etc ATP Rose Rose Hershko Ciechanover Hershko Rose Rose 3 Nature, Science, Cell March 2005 No.14 19

22 Rose Haas Wilkinson Pickart Rose 1980 Hershko/Ciechanover APF-1 Haas Wilkinson Rose Rose 3 Hershko Ciechanover Rose Hershko APC Ciechanover N-terminal ubiquitination Rose of the University of California, Irvine. Hershko found out when his cousin phoned and told him. The trio was rewarded for unravelling the mechanism behind a molecular kiss of death a tag that marks proteins for destruction. Varshavsky Hershko Varshavsky Ciechanover Rose Varshavsky Nature 431, Avram Hershko thought he might have a chance of winning this years Nobel prize for medicine. So when the results were announced last week 10 4, and his name wasn t on the list, Hershko decided to spend some time by the pool with his granddaughters: Maybe some other year, I thought While he was enjoying his holiday, Israeli radio announced on 6 October that he was a co-winner of this year s chemistry Nobel prize, together with Aaron Ciechanover, also at Haifa, and Irwin Nature 20

23 Varshavsky Hershko Varshavsky Extension Hershko Varshavsky Varshavsky Hershko Varshavsky Varshavsky N-end rule N Varshavsky Varshavsky 1984 Varshavsky Finley Ciechanover ts85 E1 Cell Cell Ubiquitin as a central cellular regulator Cell 116, S Pickart Back to the future with ubiquitin Cell 116, Varshavsky Varshavsky Daniel Finley, Stefan Jentsch, Mark Hochstrasser Varshavsky Hershko Varshavsky q1w I am sure many people are puzzled by Alex Varshavsky not being part of the prize. The way I understand it, the prize is for a ground-breaking piece of work accomplished over a short period of time that was responsible for establishing or defining the field. By those criteria Alex did not participate in the very earliest work and did not in fact begin to publish in the field until a couple of years later. q2w I am sure that you have also heard all the exciting news about the Nobel Prize to Avram, Aaron, and Ernie Rose for ubiquitination. In my view, the selection committee showed enormous wisdom in recognizing that the key early findings depended on Rose whose contributions have been widely ignored. q3w Nature In 2000, Varshavsky shared the prestigious Albert Lasker Award for Basic Medical Research with Ciechanover and Hershko for work on ubiquitin. But he is absent from the list of Nobel winners. Berg director of National Institute of General Medical Science in Bethesda suggests that this may be because Varshavsky s work, which confirmed theories about ubiquitin by studying its action in yeast, may have been judged to be closer to cell biology than biochemistry making it a step too far removed for a chemistry prize. Rose, in contrast, laid the groundwork for the 1980 papers by studying the chemistry that underlies the ubiquitin pathway. Varshavsky Varshavsky Varshavsky Varshavsky q1w I was shocked when I learned that Alex was not included. I feel very sorry for Alex. q2w I m deeply sorry for Alex, but I wish you could understand how I feel. q3w I was very disappointed to see that Alex was not recognized. I think even in the context of the Chemistry prize, his work is very important not only regarding the physiology of the pathway but also identifying the first genetically encoded degrons and the nature of the ubiquitin signal used for proteasome recognition. March 2005 No.14 21

24 Science Varshavsky Varshavsky Varshavsky Varshavsky A group of us are thinking about writing a letter to Science to highlight these important contributions of Alex. Varshavsky The idea is not to criticize the judgment of the Nobel Prize committee but to express our sympathy for Alex. This is probably much more important to me since he is my personal science hero, my mentor, and, most importantly, my friend Varshavsky Alex q1w No problem at all. Thank you anyway for sharing with us your thoughts. q2w Thank you for your thoughtful letter. I understand, and I appreciate your honesty Keystone Symposia Ubiquitin Signaling Varshavsky Varshavsky s Contributions Science 306, We suggest that the impact of Varshavsky s work on the physiology of the ubiquitin system and its relationship to fundamental processes such as mitosis and chromosome segregation justifies serious consideration for a future Nobel Prize in Physiology or Medicine. Varshavsky Baumeister, DeMartino, Deshaies, Emr, Finley, Hampton, Hochstrasser, Huber, Jentsch, Pagano, Pickart, Rechsteiner, Sommer, Tyers, Vierstra, Weissman, Wilkinson, Wolf 25 Varshavsky Alex Nature Hershko I m very, very sorry for Varshavsky. He really deserves it. The importance of what we had done only became clear after Varshavsky s work. I d be happier if there could be four winners. It must have been a very challenging political decision, because there have been huge efforts organized from some university with Alex s initiation to nominate him Varshavsky together with Hershko for various prizes, especially the Nobel Prize in Medicine. The committee clearly avoided this challenge by giving the Prize in Chemistry for the enzymology, where the discover- 22

25 Vol ies were completed before Alex even entered the field. Varshavsky Varshavsky Varshavsky Aaron Ciechanover Avram Hershko Irwin Rose 3 Hershko Nobel Lecture Banquet Speech Interview Nobel Diploma Prize Award Photo Other Resources q1w Hershko q2w 3 March 2005 No.14 23

26 HP Ciechanover Keiji! Give me this slide? 10 6 Hershko Hershko Rose Avram Hershko Interview Telephone interview with Professor Avram Hershko after the announcement of the 2004 Nobel Prize in Chemistry, October 6, Interviewer was Joanna Rose, science writer. Hello. Hello Avram. Congratulations to the prize. My name is Joanna Rose and I call from the Nobelprize.org, which is the official website of The Nobel Foundation. Yes. My congratulations to the prize. Thank you. How does it feel now? Oh, I am very happy. Very happy for my family, for my institution, my country, and for myself also. I think this is a very as you know it is a very good recognition. I m also very happy, I should add that that Irwin Rose was included, because I got many prizes before, but he was never included. And he did make a very important contribution to the discovery. So I am glad that justice was made. I really think that justice was made at this time. Did you expect the message today? No. I was out on a picnic with four granddaughters. It is a holiday today in Israel. We call it a day of a kids day. So I invited four grandchildren, and we went out for a picnic, and to a swimming pool in a kibbutz, and there I heard it from somebody heard it on the radio. I understand. But it was good. It was very exciting. Yeah. What was your first reaction when you heard it from the radio? Well, I thought I was very happy. My first reaction was I am very happy for Ernie Rose. And, also happy for myself, of course. And for Ciechanover. Can you tell me just how do you think that the Nobel Prize is going to affect your future work? I you know I enjoy bench work very much. I try to do an experiment every day, even today. And, I would like to continue with that because it s really exciting. So, I hope it won t affect too much my life. But of course you never know. There will be distractions I am sure. And there will be some duties. I sure there will be some invitations I will have to say yes to. But, more or less, I would like to continue to do my work. I think I can still contribute. Not in the same big way as twenty-five years ago, but still contributing and then still having a lot of fun at the bench. Did you realize, when you did your discovery for over twenty years ago, that it is worth a Nobel Prize? Yeah. I thought so. I wasn t waiting for it you know, but I knew already that it because the impact is really big, you know about when I started to work on ubiquitin there were about ten papers a year on ubiquitin. And now there are thousands in a year. So, it really became a kind of a cascade, and many people heard about us all over the world mind about us very big about this all over the world are working on different aspects of the ubiquitin system and different systems. So I knew it was important. But I wasn t waiting for the prize. No, I wasn t waiting for it. But of course, I am very grateful for it. I understand. Have you any good advice to young students that maybe dream about receiving the Nobel Prize in the future? Well, not about receiving the Nobel Prize, but about doing science. My advice is well that s what I did, you know, to try to find something novel, and open up new problems which is not yet reached a big level at this time, not yet interested, but you think is important. I think that s what I did about thirtyfive years ago. And then, continue with it. That s my advice. Try to find a unique problem which is important, but which is not yet in the center of the attention of biology or of chemistry. I think that is true for discoveries, that s how it should be done. So, that s my advice for young people. Yeah. My last question is, have you ever visited the Nobel website? Pardon me? Have you ever visited the Nobel website on the internet? No. Um-hmm. So, now you will be there yourself. O.K. Yes, thank you very much and have a good day. Thank you. Same to you. Thanks for calling. Bye. Bye. 24

27 Essay MBL MBL 45 MBL IT March 2005 No.14 25

28 MBL 4A 4B 26

29 P. F BP DNA Essay / NMR Kurt Wührich 2002 NMR NMR Wührich NMR IgG 15 NMR March 2005 No.14 27

30 1 NMR NMR NMR 1 100mg 3 IgG Fc Fc NMR NMR NMR NMR Ubl AR-JP 52kDa E3 1 C RING E2 N Ubl AR-JP parkin 2 RING E3 parkin Ubl 42 Ubl 2 NMR 28

31 Ubl 4 Wührich NMR NMR Ubl 3 Ubl NMR - NMR ph Ubl - Ubl NMR - Ubl - competitor 0.1mM NMR 3 10 natively unfolded protein Ubl Wührich NMR NOE A B 5 natively unfolded 800MHz NMR NOE RDC NOE Ubl RDC 2 NMR Ubl 26S Rpn10 3 intrinsic disordered region Rpn10 AR-JP Arg42 Pro 26S AR-JP 2 0.1mM NMR March 2005 No.14 29

32 NMR structural glycobiology HPLC 3 A PA 3 HPLC PA ODS ODS X Y Z HPLC HPLC WEB GALAXY 5 6 HPLC HPLC 30

33 NMR SCF Fbs1 Fbs1 7 Fbs1 SBD 10 6 N- GlcNAc 4 NMR Fbs1 SBD Fbs1-SBD NMR Fbs1 GlcNAc GlcNAc Fbs1 ERAD Fbs1 Fbs1 - Fbs1 N- PNGase PNGase SCF Fbs1 Fbs1 PNGase Fbs1 PNGase 6 NMR Fc NMR March 2005 No.14 31

34 NMR HPLC NMR 7 NMR 1. Shimura, H. et al: Nat. Genet. 25, Terreni, L. et al: Neurology 56, Sakata, E. et al: EMBO Rep. 4, Takahashi, N.: Biochem. Biophys. Res. Commun., 76, Takahashi, N., et al: Anal. Biochem., 226, N.Takahashi, N. and Kato, K.: Trends Glycosci. Glycotech., 15, Mizushima, T., et al: Nat. Struct. Mol. Biol. 11, Essay

35 T Life of Proteins No O O q1w q2w q3w q2w q3w q1w q3w q1w q3w March 2005 No.14 33

36 6 7 J

37 March 2005 No.14 35

38 5 2 q1w q2w q3w

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40 1. : Life of Proteins 13, Jacques Monod: Le hasard et la nécessité Alfred A. Knopf, Inc Douglas Adams : Don t Panic BBC Hitchhiker s Guide to the Galaxy Life of Proteins 13, Red Dwarf TV DVD PAL 8. Stephen Jay Gould : Richard Dawkins: A Devil s Chaplain 2003 pp : Life of Proteins 12, : : : Mini Reviews

41 Mini Reviews Loyola Jody Brewer XBP1: a link between the unfolded protein response, lipid biosynthesis, and biogenesis of the endoplasmic reticulum ERSE ATF6 XBP ATF XBP1 mrna IRE1 IRE1- XBP1 Ire1p-Hac1p 3 ATF6 IRE1-XBP1 Ire1p-Hac1p 1 ATF6 XBP1 ATF6 ATF6 IRE1-XBP1 IRE1 ATF6 mrna XBP1 XBP1 ERSE XBP1 XBP1 ATF6 XBP1 XBP Laurie Glimcher class II X box B XBP1 4 XBP1 DNA ATF6 Michael Green Activating Transcription Factor Ron Prywes serum response factor ATF6 March 2005 No.14 39

42 serum response Prywes ATF6 DNA PCR ATF6 site ATF6 ATF6 site UPR ATF6 site JBC 6 ATF6 site IRE1 ATF6 site ATF6 ATF6 ATF6 IRE1 IRE1 XBP1 IRE1 ERAD XBP1 ATF6 ATF6 site q2wb ATF6 DNA 2A in vitro ATF6 ATF6 site in vitro translation XBP1 XBP1 IRE1 XBP1 q1w XBP1 ATF6 site ERAD ATF6 site IRE1 EDEM IRE1-XBP1 ATF6 site 6 q2w ATF6 site UPRE JCB XBP1 UPRE Ire1p Hac1p XBP1 site Jody Linda Hendershot ATF6 Assistant Professor UPR ERSE XBP1 ERSE UPRE Glimcher XBP1 ATF6 ATF6 XBP1 UPRE Jody XBP1 NIH3T Cell ATF6 IRE1-XBP q1w 2B ERAD XBP1 40

43 C XBP XBP1 B q2w Jody mrna XBP1 XBP1 XBP1 Glimcher XBP1 XBP1 XBP1 8 XBP1 Peter Walter UPR 9 XBP1 Hac1p UPR Opi1p Hac1p 2 Opi1p Hac1p Hac1p Ire1p Hac1p UPRE 10 XBP1 Hac1p 1 HAC1 mrna XBP1 mrna IRE1 UPR UPR XBP1 UPR UPR XBP1 ATF6 ATF6 1. Sriburi R. et al., J. Cell Biol. 167, Haze K. et al., Mol. Biol. Cell 10, Yoshida H. et al.: Cell 107, Reimold A. M. et al. : Nature 412, Wang Y. et al. : J. Biol. Chem. 275, Yoshida H. et al. : Dev. Cell 4, Rush J. S. et al. : Arch. Biochem. Biophys. 284, Shaffer A.L. et al. : Immunity 21, Travers K. J. et al. : Cell 101, Ogawa N. & Mori K.: Genes Cells 9, March 2005 No.14 41

44 Mini Reviews COPII refolding q1w q2w ERAD q3w ERAD 1 3 IRE1 Inositol Requiring 1 ATF6 Activating Transcription Factor 6 eif2 PERK PKR-like ER Kinase IRE1 PERK I ATF6 II Hsp70 BiP/Grp78 1,2 BiP BiP BiP BiP BiP BiP 42

45 BiP IRE1 PERK 2 1 ATF6 BiP S1P, S2P N 3 BiP IRE1 BiP ATF6 BiP 1,4 BiP HSF1 Heat Shock Factor 1 Hsp70 HSF1 Hsp70 HSF1 3 Hsp70 BiP/Hsp70 BiP BiP BiP BiP BiP UPR Unfolded Protein Response Ire1 BiP BiP Ire1 BiP BiP UPR 5 BiP UPR D. Raden et al., 2004 Mol. Chaperone, Cold Spring Harbor Meeting Ire1 Ire1 IRE1 4 N IRE1 N N- 4 Cys 3 7 Ire N in vivo 2 N I II 130 IV III III 10 II IV Ire1 N in vitro II IV 7 in vivo Ire1 BiP BiP March 2005 No.14 43

46 adjustor 6 Prywes ATF6 BiP ATF6 BiP ATF6 BiP 8 Ire1 BiP V Ire1 BiP 2 in vivo II IV IV in vivo BiP 2 in vitro 2 native gel pull-down 7 II 2 2 BiP IV Ire1 2 II Ire1 2 V BiP 3 BiP Ire1 Ire1 BiP BiP BiP Ire1 BiP B IRE1-XBP1 PERK 1. Bertolotti, A. et al.: Nat. Cell Biol. 2, Okamura, K. et al.: Biochem. Biophys. Res. Commun. 279, Haze, K. et al.: Mol. Biol. Cell 10, Kimata, Y. et al.: Mol. Biol. Cell 14, Kohno, K. et al.: Mol. Cell. Biol. 13, Kimata, Y. et al.: J. Cell Biol. 167, Oikawa, D. et al.: submitted. 8. Shen, J. et al.: Mol. Cell. Biol. 25, Ire1 BiP Ire1 V Ire1 BiP 44

47 Mini Reviews O 1 O O O 1,2 O O protein O-mannosyltransferase; PMT PMT1 PMT7 O Pmt1p PMT PMT1 Pmt1p Pmt5p PMT2 Pmt2p Pmt3p PMT4 Pmt1p/Pmt2p Pmt5p/Pmt3p Pmt4p PMT1/PMT2 PMT4 Mnt1p Ktr1p March 2005 No.14 45

48 Ktr3p Mnn1p 4 5 1,2 1,3 110 O O O O ERAD Endoplasmic Reticulum- Associated Degradation Pmt2p O 3 O pro RNAP-I O O 3 O 4 KHN Kar2p simian virus 5 HA-neuraminidase ectodomain ERAD KHN Pmt1p Pmt2p O O KHN O KHN O 4 3 O KHN BPTI bovine pancreatic trypsin inhibitoro 5 O Pmt O 2 Pmt O O O pmt2 UPR Unfolded Protein Response UPR PMT1 PMT4 O O 3 pmt4 Ccw5p O N Ccw5p Pmt4p O N 6 3OST N Pmt4p O 46

49 pmt4 O 3 Fus1p Fus1p Pmt4p O O Fus1p pmt4 Fus1p Fus1p O Pmt1p/Pmt2p O Mid2p Fus1p O 7 Axl2/Bud10p axial budding pattern pmt4 Axl2/Bud10p N 8 Tanner 6 Axl2/Bud10p Ccw5p N ERAD PMT POMT1 POMT2 O 9,10 POMT1 POMT2 PMT4 PMT2 N- 1 O O O POMT1 11 O Pmt1p 5 SDF2-L1 stromal cell-derived factor 2-like 1 12 SDF2-L1 C HDEL SDF2-L1 BiP GRP94 PDI ERdj3p/ERj3p Hsp40 Scj1p ERdj3p/ERj3p 13 O O PMT O O O pro 1. Strahl-Bolsinger, S., Gentzsch, M., Tanner, W. : Biochim. Biophys. Acta. 1426, Willer, T. et al.: Curr. Opin. Struct. Biol. 13, Nakatsukasa, K. et al.: J. Biol. Chem. 279, Vashist, S. et al.: J. Cell Biol. 155, Coughlan, C.M. et al.: J. Biol. Chem. 279, Ecker, M. et al.: EMBO Rep. 4, Proszynski, T.J., Simons, K., Bagnat, M.: Mol. Biol. Cell 15, Sanders, S.L., Gentzsch, M., Tanner, W., Herskowitz, I.: J. Cell Biol. 145, Manya, H. et al.: Proc. Natl. Acad. Sci. U.S.A. 101, Ichimiya, T. et al. : J. Biol. Chem. 279, Willer, T. et al.: Proc. Natl. Acad. Sci. U.S.A. 101, Fukuda, S. et al.: Biochem. Biophys. Res. Commun. 280, Bies, C. et al.: Biol. Chem. 385, March 2005 No.14 47

50 Mini Reviews E E E RseA RseA DegS RseP YaeL a 2 E 1 1,2 ER ATF6 2 2 RseP S2P RIP: Regulated Intramembrane Proteolysis 3 E RseP ATF6 ER S1P/S2P E RseA DegS 4,5 DegS PDZ DegS C --YYF-COOH DegS PDZ RseA DegS RseP DegS ON/OFF DegS PDZ PDZ = DegS PDZ DegS 48

51 PDZ DegS RseA E RseA E E RseP RseA ClpXP RseA E 6 clpxp E ClpXP E RseP DegS RseA RseP RseA DegS RseA RseP PDZ 7 RseA 2 Gln Q1 Q2 Gln Ala DegS RseP RseP PDZ RseA PDZ RseA Q1/Q2 RseP PDZ RseA RseB RseP RseA 8 RseB RseA RseB RseA RseB RseA E RseP RseA 2 RseP rsea rseb rsep rse yael 4 degp clp in vitro 1 fts fts fts1 2 ftsi ftsh fts 95 1 March 2005 No.14 49

52 RseA RseP 2 RseA MBP MBP-RseA140 2 DegS RseA DegS RseP MBP 9 2 RseA Ala108 Cys109 RseP MBP- RseA140 in vitro RseP in vivo S2P 9 RseP RseA RseP DegS RseA E 1,2 RseP RseA RseP LacY RseA in vivo in vitro 9 RseP LacYTM1 TM Pro 9 RseP RIP SPP RIP RseP RseA RseP - SPP SPP RseP SPP rsep ATP FtsH FtsH RseP FtsH RseP SPP RseP 10 RIP -- RIP S2P SREBP S2P RseP PDZ RseA RIP intramembrane 50

53 RIP 1. Ades, S. E.: Curr. Opin. Microbiol. 7, ) 2. Alba, B. M., and Gross, C. A. : Mol. Microbiol. 52, : 48, Walsh, N. P., Alba, B. M., Bose, B., Gross, C. A., and Sauer, R. T.: Cell 113, Wilken, C., Kitzing, K., Kurzbauer, R., Ehrmann, M., and Clausen, T.: Cell 117, Flynn, J. M., Levchenko, I., Sauer, R. T., and Baker, T. A.: Genes Dev. 18, Kanehara, K., Ito, K., and Akiyama, Y.: EMBO J. 22, Grigorova, I. L., Chaba, R., Zhong, H. J., Alba, B. M., Rhodius, V., Herman, C., and Gross, C. A. : Genes Dev. 18, Akiyama, Y., Kanehara, K., and Ito, K.: EMBO J. 23, Meeting Report m March 2005 No.14 51

54 ATF QBP KaiA KaiB KaiC KaiA C 2 N 24 KaiA KaiC ATP KaiC-ATP-KaiA GroES X SpoU SpoU 6M 1 7 N C Histag C Trp FRET PURE system DnaK, DnaJ GroEL mrna- -GroEL GroEL co-translational ER HSP47 HSP47 Arg HSP47 Low Affinity Binding Site Hsp57 Sup35, PolyQ HSP104 RNQ1 Sup35 52

55 PSI + PSI - Hsp104 PSI + HSP104 ClpB 2 Sup35 HSP104 HSP104 ATP Sup March 2005 No.14 53

56 ITC 1 100nm 10 M 1 GroEL GroES Football MKKS MKKSP MKKSP CCT CCT 8 Ring GFP MKKSP FRAP FLIP MKKSP P48 RNA P36 P36 N C P48 P36 P48 RNA shsp CCT ADP-AlFx Thermosome ADP-BeFx Thermosome CCT ATP ATP shsp shsp shsp HSP 54

57 Meeting Report March 2005 No.14 55

58 N immobile hop-diffusion terminal misfolding N cre/loxp ADAMs calr3 - -calr3 - - KDEL KDEL BiP KDEL BiP KDEL CFTR PLD 56

59 CFTR curcumin CFTR trafficking accelerator Ryh1 GTP Plc1 C Ryh1 Ryh1 Plc March 2005 No.14 57

60 Plc1 v-snare Rab G Rab13 JRAB TJ JRAB TJ TJ TJ Rab13-JRAB t-snare Psy1 Psy1 FM Myo1 Sec9 Psy1 t-snare Psy1 Psy1 1 A SLO A 10 ATG5 A importin hsc70 importin transportin hsc70 hsc70 importin transportin importin transportin 58

61 SUMO SUMO SUMO SUMO SUMO E3 RanBP2 RING HECT RanBP2 SUMO p97/vcp ERAD p97/vcp AAA NVL2 RNA 60S NVL2 L5 DNA RNA RNA NP RAF-2 RAF-2 p48/uap56 NP RNP p36 p48 NP RNA DNA- 4 4 March 2005 No.14 59

62 Meeting Report ADAM, N C RFP GFP Sec61 Sec61, ERAD 60

63 Sec61 Sec61 Sec61 Sec61, ERAD Sec61, Atg8 LC3 GFP-LC3 A Apg5 MKK MKKSP MKKSP CCT MKK MKKSP MKKS 4 MKKSP MKK MKKSP MKKSP MAP DYRK DYRK DYRK Hsp90 Hsp90 DYRK DYRK March 2005 No.14 61

64 Lys48, 2 2, Herp Herp N Herp Herp NHK Herp ERAD Herp SCF Fbsl Fbs1 SCF Fbsl ERAD VCP VCP ATPase SCF Fbsl ERAD SCF Fbsl Fbs1, Fbsl X

65 ERAD 3 Fbsl IRP2 E3 HOIL-1 HOIP HOIP RING-finger UBA Zinc-finger UBA HOIL-1N HOIP HOIL-1 B HBx 3 HBx NF-kB HOIP UBA RING-finger NF-kB HOIP HOIL-1 IPR2, PKC HBx, E March 2005 No.14 63

66 (HOlL-1) Culin2 Cullin5 E3 VHL Cul2-box Cul2/Rbx1 Cul2 VHL VHL-box SOCS-box C Cul5-box Cul5/Rbx2 VHL Cul5-box Cul5/Rbx2 SOCS-box Cul2-box Cul2/Rbx1 E3 E3 A XDRP1 XDRP1 Dsk2 Sem1 Sem1 26S Rpn10 26S Sem1 Sem1 Dsk2 A Rpn10c BAG Scythe Scythe N EMBO J J. Cell Biol J. Biol. Chem Nature

67 Scythe Scythe SUMO, SUMO SUMO E1 E2 SUMO SUMO SUMO-2 Chaps SUMO-2 SUMO in situ DNA SUMO 5 SUMO SUMO E2 ubc9 ubc9-1 SUMO E2 ubc9 ubc9-1 G2 S DNA SUMO AAA AAA ATP FtsH FtsH ATPase Arg ATPase Walker B Arg ATPase C. elegans AAA AAA 26S ATPase Meeting Report M1???! March 2005 No.14 65

68 M1!! 4 Drp1!! 66

69 Meeting Report The 1st Pacific-Rim International Conference on Protein Science 4 Protein Society Protein Society Protein Society Symposium 10 Eisenberg Protein Society Meeting 2001 Chaperone Newsletter, No.9, p Christopher Dobson Cambridge Sung-Hou Kim Plenary Lectures JR Plenary Lecture Dobson NMR Post Evolutional Disease March 2005 No.14 67

70 in vivo Folding, Transport, and Quality Control 5 Sup35 20 ClpB Hsoumin Li Tic40 Hsp70 NMR local unfolding global unfolding EDEM ER Asn-X-Ser Thr Asn in vitro EDEM RIP YaeL 5 Protein Society Robert Matthews 30 Rossmann fold / / / TIM GroEL 68

71 N C Richard Simpson 4 / 3 Molecular strategy for designing robust protein George Makhatadze 3000 Protein Society Meeting March 2005 No.14 69

72 Meeting Report Cold Spring Harbor Laboratory Meeting DsbA-DsbB- UQ Jim Bardwell Michigan Univ. CSHL meeting 2 Bardwell Glockshuber Dsb EMBO J. J. Biol. Chem. PNAS Bardwell 2 J. Biol. Chem. 277, ; PNAS 100, A4 5 3 Bardwell DsbB DsbA-DsbB-UQ CSHL meeting DsbB Ero1p from yeast Kaiser MIT Fass Weizman Inst. DsbB Ero1p DsbB DsbB Ero1p FAD Ero1p May 28 Cell N C C 100 PDI DTT UCSF Weissman Ero1p FAD FAD Weissman Ero1p something missing Ero1p 70

73 Unfolded Protein Response Ero1 David Ron truncated Cell CSHL meeting Rick Morimoto opening remark C. elegans poly-q cell toxicity poly-q aggregate HSF-1 chaperone network Susan Lindquist Hsp104 Sup35 sup35 ATP vitro PSI + Hsp104 Sup35 Weissman Sup35 psi - psi - PSI + Sup35 T m 2 Kopito Stanford Univ. O Halloran Northwestern Univ. superoxide disumutase-1 SOD1 ALS SOD1 SOD1 S-S S-S CCS SOD1 S-S S-S SOD1 Kenyon UCSF aging heatshock transcription factor HSF-1 DAF-2, DAF-16 Craig Wisconsin Univ. Ribosome-associated Hsp70s, Ssb & Ssz1 J-type Zuo1 exit tunnel Ssb nascent chain Ssz1 Zuo1 Craig Zuo1 Ssb ATPase Ssz1-Zuo 1 Ssb Ssz1 Zuo1 J-type MIT Sauer ClpXP substrate recognition, unfolding & degradation Sauer Matouschek Northwestern Univ. Clp DHFR, Barnase NF- B March 2005 No.14 71

74 Cubitus interruptus Vigh Gdansk Univ small Hsps, IbpA & IbpB Bardwell Dsb TAT motility 2S-2Fe Science S-S Dsb system characterize CSHL meeting D3 T.A. Baker J. M. Flynn SspB ClpXP RseA N SspB SsrA tag ClpXP RseA E anti- DegS YaeL RseA anti- YaeL RseA N N-RseA DegS YaeL SspB N-RseA ClpP trap ClpP in vitro N-RseA ClpXP SspB clpx sspb ClpXP N-RseA RseA E DegS YaeL ClpXP R. Kopito Atg5 Atg12 co-localize Atg8 Atg12 sirna T. J. Silhavy D. D. Isaac Cpx Cpx 2 CpxA/CpxR CpxP CpxA ( ) CpxP DegP DegP cpxp CpxP CpxP DegP New York 72

75 Meeting Report 004 Cold Spring Harbor Laboratory Molecular Chaperones and the Heat Shock Response meeting CHSL meeting 2 Yale Horwich 2 Horwich 2 2 Horwich SDS- PAGE Horwich GroEL GroEL Lu Pin1 Ser/Thr-Pro PPIase Pin1 Pin1 Pin1 KO tau Lu March 2005 No.14 73

76 Zoghbi Ataxin1 polyq Ataxin CHIP Ataxin CHIP Hsp90 EMBO Hsp90 symposium Hsp90 Hsp90 Geldanamycin Hsp90 HSF small HSP shsp Hsp70 buffer Kenyon 2 daf-2 daf-2 shsp Vigh shsp Weiss shsp transgenic tomato shsp shsp GM Buchner shsp shsp N 21 shsp shsp shsp Bukau Ludlam Trigger factor TF 50S TF N TF TF TF ribosome Nature Hartl -gal post translational co-translational post translational TF, DnaK TF, DnaK co-translational co-translational Hartl GroEL Lindquist Hsp104 sup35nm Hsp104 ATP Science in vivo Hsp104 Lindquist vs Lindquist 74

77 Weisman 2 Weisman FASEB meeting Weisman Weisman Tkach Hsp104 N 17 Tsai T. thermophilus ClpB AMP-PNP, ADP, ADP AlFx ClpB ClpB Hsp104 ClpB coiled-coil Horwich Tsai Tsai Acknowledge Bukau Lindquist ClpB AAA Bukau Tsai Bukau TF TF ClpB Hsp90-p50 SecA SecA/SecB PI PI Yale March 2005 No.14 75

78 Meeting Report The Role of Ubiquitylation in the DNA Repair Pathway DNA Life of Proteins DNA DNA DNA DDB1-DDB2/XPE E3 XPC DDB1-CSA E3 3 Dr. Weidong Wang NIH FANCD2 4 DNA Dr. Stefan Jentsch Max Planck Institute DNA PCNA Stefan Jentsch DNA SUMO PCNA 164 Rad6, Rad18 error-prone repair PCNA Mms2, Ubc13, Rad5 error-free repair Jentsch 2 76

79 Journal of Cell Biology 7 5 D 2 FANCD2E3 5 E3 Wang A FAAP250, 100, 95, 43 FAAP43 E3 PHD finger E3 L FAAP43/FANCL/PHF9 FANCD2 E3 Nat. Genet. 35, FAAP95 FAAP250 FAAP95 B FAAP250 I J A B C E F G L E3 D2 Nature 419, DNA E2 Ubc7 mat1 locus pop-out rdna Ubc7 ubc7 UV MMS DNA E3 Pcu3 Cullin3 UV MMS Rhp51 Rad51 Ubc7-Pcu3 DNA DNA Weidong Wang Cullin4A E3 XP CS DNA A CSA E DDB2/XPE Cullin4A- DDB1 E3 CSA DDB2/XPE E3 E3 SCF CSA DDB2/XPE Cullin4A-DDB1 E3 Cell 113, XP-E CSA E3 CSA DDB2/XPE CSA RNA pol II DDB2/XPE C XPC in vivo, in vitro DDB2/XPE Cullin4A-DDB1-DDB2/XPE March 2005 No.14 77

80 XPE E3 XP XPC The Role of Ubiquitylation in the DNA Repair Pathway DNA DNA E A Dr. Wang NIH , , , 23, Meeting Report FASEB 2 Stanley Prusiner UCSF 78

81 2 3 3 Jonathan Weissman UCSF Weissman Weissman Sup35 Sup35 Sup35 in vivo Hsp104 Sup35 PSI + Hsp104 Hsp104 Sup35 Sup35 Hsp104 Shorter 1 in vivo Hsp104 Hsp104 Sup35 Hsp104 Sup35 Hsp104 Sup35 Hsp104 5 Cold Spring Harbor Meeting Weissman 5 FASEB Weissman Sup35 Hsp104 Inoue, Kishimoto 2 Weissman Sup35 Cy5 Sup35 Cy3 Sup35 Cy Sup35 2 Sup Sup35 3 March 2005 No.14 79

82 X X Sup ~10 artifact Perutz 4 NM Stanley Prusiner UCSF 2 TEM -helix 3 Ronald Wetzel Tennessee Univ. A N 1-14 C Wetzel cross-seeding cross-seeding Robert Tycko National Institutes of Health A 1-40 Agitated non-move 13 C 13 C- 13 C NMR Agitated not-move parallel D23 K28 Agitated 0.35 nm not-move 0.45 nm 0.1 nm Agitated 80

83 A 1-40 Agitated artifact 2 Wetzel A Tycko A D23 K28 Wetzel massper-length MPLA X Louise C Serpell Cambridge Univ. Sup35 Sup35 ph 6.8 AFM , TEM ~120 X HEPES ph 7.5 X Islet Amyloid Polypeptide IAPP KFFEAAAKKFFE TEM X X Serpell Wetzel Serpell semi-dry Serpell Serpell X Serpell Ure2 TEM Ure2 Tris-HCl ph AFM TEM AFM TEM TEM ~ AFM TEM March 2005 No.14 81

84 TEM Helen R. Saibil Birkbeck College London Ronald Melki CNRS Ure2 FTIR X X Serpell Ulrich Baxa National Institutes of Health Ure strand proteinase K Melki Ure2 Ure2 David Eisenberg UCLA RNase A dimer 3 poly-q poly-q X cross RNase A N A19 19 A C H H RNase A-poly-Q RNase A RNase A 1. Shorter J, Lindquist S. : Science 304, Inoue Y., Kishimoto A., Hirao J., Yoshida M., Taguchi H.: J. Biol. Chem. 276,

85 3. Kishimoto A., Hasegawa K., Suzuki H., Taguchi H., Namba K., Yoshida M.: Biochem. Biophys. Res. Commun. 315, Perutz M.F., Finch J.T., Berriman J., Lesk A.: Proc. Natl. Acad. Sci. U S A 99, Meeting Report International Congress on Stress Responses in Biology and Medicine 1 100km/h 1 Chateau Mont Sainte-Anne 1 2 Cell Stress Society International CSSI North American Hyperthermia Society NAHS March 2005 No.14 83

86 2 Cell Death and Survival Strategies D. Mosser Guelph J. Landry Quebec Hsp S. Borkan Boston Hsp70 Hsp72 Bcl-2 D. Mosser Guelph Hsp70 pro-apoptotic Bax Bax Hsp70 ATPase domain JNK C. Garrido Dijon Apaf1-KO Hsp70 Apoptosis inducing factor AIF AIF caspase Hsp70 AIF AIF Hsp70 AIF Hsp70 M. Jäättela Copenhagen Hsp70 lysosome Hsp Hsp70 cathepsin Hsp70 D. Green Hsp70 R. Morimoto Evanston C. elegans HSF-1 polyq Hsp70 polyq aggregate HSF-1 polyq polyq rescue 220 Hsp70 Hsp60 Hsp40 E1 E3 E2 insulin-like signaling pathway ILS mutant C. elegans HSF-1 hsf-1 RNAi Hsp Hsp ILS Hsp Hsp 9 11 Stress, Aging and Degenarative Diseases G. Boulianne R. WadhwaG. Boulianne 84

87 Tronto Drosophila motor neuron specific SOD1 DNA array SOD R.W. Tanguay Quebec G. Lithgow Buck Institute small heat shock protein Chaperone and cellular signaling J. Huot M. Sherman J. Huot Quebec introduction Hsp90- Hsp70-Hsp27 Hsp90 src kinase Hsp70 HSF-1 Hsp27 MAP kinase R. Morimoto Evanston Hsp70 Bag1 Bag1 Raf-1 Bag1 Hsp70 Hsp70 Hsp70 Bag1 Raf- 1 Hsp70-Bag1 Ask1 TNF-a MAPKKK E.J. Choi Seoul H2O2 Hsp70 Ask1 MAPKK Hsp70 ATPase domain Hsp90 L. Neckers NCI, Rockville Hsp90 ATP ErbB2 Hsp90 ErbB2 CHIP Hsp70 Hsp90 D. Smith Scotsdale, Arizona Hsp90 immunophilin J. Yaglom Boston UnivHS Hsp70 MAP kinase HS MAP kinase JNK HS MAP kinase ERK1 ERK2 ERK-activating kinase MEK1/2 ERK MKP-3 HS MEK Hsp70 MKP-3 ERK MEK W. Van. Molle Zwijnaarde, Belgium TNFa Hsp70 Hsp70 Hsp70 Hsp70 Hsp70 Hsp70 Hsp70 IL-6 NO Hsp70 Avrom Caplan Mount Sinai NY v-src folding Cdc37 Cdc37 Hsp90 Hop code STI1 Hsp90 v-src v-src Cdc37 STI1 v-src Cdc37 Hsp90 M. Sherman Boston Univ Hsp70 Hsp70 ERK Hsp March 2005 No.14 85

88 Meeting Report ClpB-DnaK ClpB DnaK-DnaJ-DafA DafA DnaK-DnaJ ClpB DnaK DnaJ 86

89 DafA DnaK-DnaJ-DafA DafA DnaK DnaJ ClpB ClpB Hsp104 Sup35 Sup35 ATP 2 2m Thioflavin T 2m Anfinsen 1962 TTR TTR NMR 15nm TTR QBP1 in vitro in vivo QBP1 11 QBP1 24,000 TOM40 Hsp70 in vivo in vitro in silico in silico CASP CASP NMR Thermus thermophilus E. coli GroEL/GroES GroEL GroES E. coli GroEL March 2005 No.14 87

90 3 RISM GFP GFP in vitro ,000 Meeting Report Plenary Lecture James Rothman Columbia Univ. Arthur Johnson Texas A&M Univ. Peter Rehling Univ. Freiburg Wolf Kunau Univ. Bochum 3 Arthur Johnson Texas A&M Univ. BiP BiP ATP BiP collisional quenching BiP ATP 88

91 ATP BiP 2 COPII 4 GTPase Sar1p Sar1p GTP FRET Sec23/24p Bet1p FRET GDP GTP Export Sar1p GTP Sar1p GTP Sar1p GTP/GDP Sec12p GTP GFP RFP importin importin importin GTPase Ran dominant negative importin UV importin Ran ATP Ran ATP GTP GTP Ran GTP Ran importin 4 Peter Rehling Univ. Freiburg Tim23 Protein A Tim Tim17, Tim23, Tim50 Tim44, mhsp70 Pam presequence translocase associated motor 16 =Tim16 Pam18 =Tim14 mhsp70 ATPase Pam18 J ATPase mhsp70 Tim44 Pam16 Pam18 ATPase Pam18 Tim Pam18 Pam16 Pam18 Tim21 Tim23-Protein A Tim21 Tim23 BlueNative PAGE core Tim23* Tim21 Tim23* Tim21 Tim23* Tim21 Tim23 GTPase Mitofusin1 Mitofusin2 Mitofusin1 GTP tethering Mitofusin1 55kDa Dynamin March 2005 No.14 89

92 OPA1 N m-aaa q1waaa Pex1/Pex6 Pex26 Pex1 Pex6 Pex1/Pex6 Pex26 q2w Pex5 Pex5 in vitro Pex5 ATP Pex5 Pex14 import Pex1/Pex6/pex26 export q3w Pex14 Pex14 Pex5 Pex13 Pex19 Pex14 Pex13 Pex19 Pex5 import Pex14 q4wpex19 PMP Pex19 Pex3 Pex19 PMP q5wdynamin Drp1/Dlp1 Drp1 GTPase Drp1 Wolf H. Kunau Univ. Bochum PTS1 Pex5 long isoform Pex5pL PTS2 Pex7 PTS2 Pex5 PTS1 Pex5 Pex7 Pex18 Pex7 Pex5 PTS1 C PTS1 N Fox1p N Pex18 Pex7 Pex5 PTS1 PTS1 Pex18 Pex5 N Pex7 PTS2 Meeting Report SCF Fbx2 Fbs1 Fbs1 Fbx2 NMR & X Fbs1 Fbs1 Fbox Skp1, Cul1 E3 Fbs1 90

93 pull-down assay Fbs1 ERAD Fbs1 ERAD Fbs1 ERAD Fbs1 Fbs1 Fbs2 N- PNGase N- ERAD PNGase PNG1 PNG1 ERAD PNGase DNA Rad23 PNGase-Rad23 MS PNGase PNGase ENGase Man5 E- Man5-9 more important than you think N- NMR VIP36 Fbs Fbs1 Man 3 GlcNAc 2 N- ER N-PNGase Fbs1 PNGase VIP36 ER apical NMR VIP36 A- Man 1 2Man 1 2Man 3 VIP36 ph (ph5.5) VIP VIP36 3 VIP36 NMR VIP36 (FAC) FAC SPR FAC FAC March 2005 No.14 91

94 FAC N- ERAD ER Man 9 Man 8 B ERAD EDEM Man 8 B Man 9 Man 8 B I ER-ManI EDEM ERAD ERAD 1-antitripsin NHK ERAD NHK ER-ManI Glc Man 9 Glc Man 8 EDEM ERAD EDEM folding 1-antitrypsin NHK EDEM soledem ERAD UPR up-regulation Northern in situ hybridization EDEM EDEM GGT GGT II N CT CT FRAP FRET GGT GGT Unfolded protein response PR ER 14 Glc 3 Man 9 GlcNAc 2 lipid-linked oligosaccharides; LLO ER LLO UPR LLO UPR UPR ER LLO LLO 6 UPR up-regulation 6 UPR LLO LLO 3 UPR LLO ALG12 upregulation 92

95 UPR 5-15min ER 30-60min PERK LLO 3-5 h LLO UPR UPR 3 Glycogene project Glycogene project MS/MS/MS MS3 Glycobiology 3 Meeting Report GroEL/ES GFP 3 March 2005 No.14 93

96 Thermus GroEL/ES GroEL/ES ClpB Ying Beiwen GroEL/ES co-translational metk mrna GroEL PURE DsbA DsbB LolCDE ATP ATP LolA Blue native PAGE CRL CRL I PSI-B PSI-E RuBisCO 94

97 Meeting Report Endo, Neupert, Pfanner 3 3 Tom13 Tom38 Tom13 SAM TOM40 Tom38 Tom40 SAM TIM22 TIM23 -DHFR 6 TIM22 2 TIM22 TIM23 mthsp70 Tom40 Tom40 E. coli 7030 March 2005 No.14 95

98 E E E DegS E RseA RseP RseA E RseB DegS E RseP RseA DegS RseP RseA E in vitro Gross RseP Site II UPR Gross Regulated intramembrane proteolysis Rip Rip ATF6 S-S ATF6 tunicamycin Site I S1P ATF6 LIP S1P ATF6 ATF6 ATF6 S1P ATF6 UPR ATF6 Parkin Parkin synuclein in vitro in vivo synuclein -synuclein 96

99 Meeting Report AAA+ 1 AAA+ 6 RuvB Arginine174 ATP p97-mediated membrane fusion VCP ClpB AAA Pex1p NMR AAA FtsH AAA+ TypeIII AAA ATPases associated with diverse cellular activities 20 ATPase AAA ATP March 2005 No.14 97

100 6 ATP Walker A Walker B ATP fidgetin RuvB ATP ATPase ATP Pex1p Pex6p AAA Pex26p Pex5p Pex1p 6 Pex6p Pex26p ATP ATP AAA N p97 VCP Cdc48p p47 VCIP135 p47 p47 p47 p47 p37 p97-p47 syntaxin5 p97-p37 SNARE p97 VCP ATPase Pex1p Skd1 N NMR ClpB ClpB Hsp104 ClpB 98

101 Hsp104 Hsp104 VCP AAA+ ClpXP AAA+ N C AAA FtsH 3 2 Daniel N. Hebert N-linked glycans act as protein maturation and quality control tags in the endoplasmic reticulum 2 -Mannosidase-like proteins involved in glycoprotein ERAD ER ManI EDEM ER degradation enhancing -mannosidase-like protein ERAD EDEM N ERAD NHK EDEM SolE: soluble homologue of EDEM EDEM3 * SolE -mannosidase C KDEL EDEM SolE EDEM SolE nerve ganglia cerebellum SolE EDEM NHK NHK-QQQ NHK EDEM EDEM2 * NHK Pathological roles of the ER stress-chop pathway 3 PERK, ATF6, Ire1 CHOP 3 CHOP Bax CHOP Hsp70-DnaJ Bax CHOP CHOP Bax CHOP 4 Ron Kopito Microtubule-dependent autophagy is required for cellular clearance of aggregated huntingtin - March 2005 No.14 99

102 Atg8, Atg12 Atg16 RNA 2 Mammalian E4 is required for prevention of neuronal degeneration E1, E2, E3 3 E4 SCA3 MJD ataxin-3 E4 E4B VCP p97 Cdc48 AAA ataxin-3 E4B C U box E4B ataxin-3 E4B dominant-negative U box ataxin-3 E4B E4B E4B E4B SCA3 * EDEM2 web site soluble EDEM EDEM3 EDEM EDEM2 publish 100

103 Meeting Report / / IICBGlc ptsg mrna RNase E RNA SgrS RNA Hfq non-coding RNA ptsg mrna IICBGlc IICBGlc ptsg mrna SgrS RNA RNaseE March 2005 No

104 non-coding RNA mrna 2 SecA ATPase q1wsecm seca SecM SecM SecA SecA SecA q2w SecA ATP post structure 3 q1w PERK ATF6 IRE1 ATF6 IRE1 IRE1 XBP1 pre-mrna ATF6 ATF6 q2watf6 IRE1 ATF6 IRE1 IRE1 BiP ERAD EDEM 4 RseP YaeLRseP RIP: regulated intramembrane proteolysis RseA RseP in vivo in vitro RseP RseA helix-destabilizing residue RseP 5 N Fbs1 SCF Fbs ERAD p97 ERAD Fbs1 p97 Fbs1 1 p97 ATP Fbs1 Fbs1 Fbs1 SCF Fbs ERAD Fbs1 ERAD 6 COPII COPII GTPase Sar1p FRET COPII Sar1p GTP Sar1p GTP Sar1p 102

105 Sar1p GTP Sar1 Atg5 Replay March 2005 No

106

107 March 2005 No

108 Calendar Zakopane Maciej Zylicz and Ulrich F. Hartl Protein folding, misfolding and aggregation / Folding in Mitochondria and ER / Chaperones in protein degradation / Misfolding diseases / Molecular chaperones and cancer Ineke Braakman, Johannes Buchner, Bernd Bukau, Aaron Ciechanover, Elizabeth Craig, Peter Csermely, Douglas Cyr, Christopher M. Dobson, Judith Frydman, Carmen Carrido, Costa Georgopoulos, F. Ulrich Hartl, Ari Helenius, Jörg Höhfeld, Arthur Horwich, Ted Hupp, Marja Jäättelä, Harm H. Kampinga, Thomas Langer, Krzysztof Liberek, Ronald Melki, Richard Morimoto, Gabriele Multhoff, Kazuhiro Nagata, Walter Neupert, Didier Picard, Giorgio Parmiani, Helen Saibil, Mick Tuite, Georg Wick, Paul Workman, Rolf D. Issels WWW Colby-Sawyer College Harris D. Bernstein, Ross E. Dalbey, and Jugen Soll Roland Beckmann, Ralph Henry, Olaf Scheneewind, Toshiya Endo, Carla Koehler, Klaus Pfanner, Rosemary Stuart, Arnold Driessen, Matthis Muller, Tracy Palmer, Manu Hegde, Hidde Ploegh, Yihong Ye, Ralf Erdmann, Yukio Fujiki, Steven Gould, Peter Christie, Vassilis Koronakis, Tony Pugsley, Jan Tommassen, Ken Cline, Danny Schnell, Steve Theg, Bill Dowhan, Art Johnson, Andreas Kuhn, Bill Skach, Steve Dowdy, Francoise Jacob-Dubbuisson, Anna Rubartelli WWW Salve Regina University Dennis J. Thiele, Peter Walter Elizabeth Craig, Bernd Bukau, Carol Gross, David Ron, Jonathan Weissman, Jeffrey Brodsky, Lea Sistonen, Akira Nakai, Xinnian Dong, Michele Toledano, Richard Morimoto, John Lis, Caroline Jolly, James Goodrich, Ivor Benjamin, Jeffrey Robbins, Kevin Morano, Doug Green, ROn Kopito, Tso-Pang Yao, Cynthia Kenyon, Lenny Guarentre, Susan Lindquist, Liming Li WWW Tomar Cell Stress Society International ( Claudia Rodrigues-Pousada Aaron Ciechanover WWW 106

109 Epilogue 1 1 2

110 Life of Proteins Chaperone Newsletter