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Altered cofactor regulation with disease-associated p97/VCP mutations

Zhang, Xiaoyi and Gui, Lin and Zhang, Xiaoyan and Bulfer, Stacie L. and Sanghez, Valentina and Wong, Daniel E. and Lee, YouJin and Lehmann, Lynn and Lee, James Siho and Shih, Pei-Yin and Lin, Henry J. and Iacovino, Michelina and Weihl, Conrad C. and Arkin, Michelle R. and Wang, Yanzhuang and Chou, Tsui-Fen (2015) Altered cofactor regulation with disease-associated p97/VCP mutations. Proceedings of the National Academy of Sciences of the United States of America, 112 (14). E1705-E1714. ISSN 0027-8424. PMCID PMC4394316. https://resolver.caltech.edu/CaltechAUTHORS:20150318-093025125

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Abstract

Dominant mutations in p97/VCP (valosin-containing protein) cause a rare multisystem degenerative disease with varied phenotypes that include inclusion body myopathy, Paget’s disease of bone, frontotemporal dementia, and amyotrophic lateral sclerosis. p97 disease mutants have altered N-domain conformations, elevated ATPase activity, and altered cofactor association. We have now discovered a previously unidentified disease-relevant functional property of p97 by identifying how the cofactors p37 and p47 regulate p97 ATPase activity. We define p37 as, to our knowledge, the first known p97-activating cofactor, which enhances the catalytic efficiency (k_(cat)/K_m) of p97 by 11-fold. Whereas both p37 and p47 decrease the K_m of ATP in p97, p37 increases the k_(cat) of p97. In contrast, regulation by p47 is biphasic, with decreased k_(cat) at low levels but increased k_(cat) at higher levels. By deleting a region of p47 that lacks homology to p37 (amino acids 69–92), we changed p47 from an inhibitory cofactor to an activating cofactor, similar to p37. Our data suggest that cofactors regulate p97 ATPase activity by binding to the N domain. Induced conformation changes affect ADP/ATP binding at the D1 domain, which in turn controls ATPase cycling. Most importantly, we found that the D2 domain of disease mutants failed to be activated by p37 or p47. Our results show that cofactors play a critical role in controlling p97 ATPase activity, and suggest that lack of cofactor-regulated communication may contribute to p97-associated disease pathogenesis.


Item Type:Article
Related URLs:
URLURL TypeDescription
http://dx.doi.org/10.1073/pnas.1418820112 DOIArticle
http://www.pnas.org/content/112/14/E1705PublisherArticle
http://www.pnas.org/lookup/suppl/doi:10.1073/pnas.1418820112/-/DCSupplementalPublisherSupporting Information
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4394316/PubMed CentralArticle
ORCID:
AuthorORCID
Sanghez, Valentina0000-0002-4131-7480
Shih, Pei-Yin0000-0003-3082-9242
Iacovino, Michelina0000-0002-1076-3146
Wang, Yanzhuang0000-0002-1864-7094
Chou, Tsui-Fen0000-0003-2410-2186
Additional Information:© 2015 National Academy of Sciences. Freely available online through the PNAS open access option. Edited by William J. Lennarz, Stony Brook University, Stony Brook, NY, and approved February 10, 2015 (received for review September 30, 2014) Published online before print March 16, 2015, doi: 10.1073/pnas.1418820112. T.-F.C. was in part supported by the National Center for Advancing Translational Sciences through UCLA Clinical Translational Science Institute (CTSI) Grant UL1TR000124 and the LA BioMed Seed Grant Program (20826-01). S.L.B. and M.R.A. were funded in part with federal funds from the National Cancer Institute, National Institutes of Health, under Contract HHSN261200800001E, through the NExT Chemical Biology Consortium. (The content of this publication does not necessarily reflect the views or policies of the Department of Health and Human Services, nor does mention of trade names, commercial products, or organizations imply endorsement by the US Government.) Y.W. is supported in part by the National Institutes of Health (Grants GM087364 and GM105920), American Cancer Society (Grant RGS-09-278-01-CSM), MCubed, and the Fastforward Protein Folding Disease Initiative of The University of Michigan. C.C.W. is supported in part by the National Institutes of Health (Grants AG031867 and AG042095). T.-F.C. is a member of the UCLA Jonsson Comprehensive Cancer Center. Author contributions: Xiaoyi Zhang, L.G., Xiaoyan Zhang, S.L.B., Y.L., C.C.W., Y.W., and T.-F.C. designed research; Xiaoyi Zhang, L.G., Xiaoyan Zhang, S.L.B., D.E.W., Y.L., and L.L. performed research; V.S., C.C.W., Y.W., and T.-F.C. contributed new reagents/analytic tools; Xiaoyi Zhang, L.G., Xiaoyan Zhang, S.L.B., V.S., D.E.W., Y.L., L.L., J.S.L., P.-Y.S., H.J.L., M.I., C.C.W., M.R.A., Y.W., and T.-F.C. analyzed data; and S.L.B., D.E.W., J.S.L., P.-Y.S., H.J.L., M.I., C.C.W., M.R.A., Y.W., and T.-F.C. wrote the paper. The authors declare no conflict of interest. This article is a PNAS Direct Submission. This article contains supporting information online at www.pnas.org/lookup/suppl/doi:10.1073/pnas.1418820112/-/DCSupplemental.
Funders:
Funding AgencyGrant Number
NIHUL1TR000124
LA BioMed Seed Grant program20826-01
NIHHHSN261200800001E
NExT Chemical Biology ConsortiumUNSPECIFIED
NIHGM087364
NIHGM105920
American Cancer SocietyRGS-09-278-01-CSM
MCubedUNSPECIFIED
Fastforward Protein Folding Disease Initiative, University of MichiganUNSPECIFIED
NIHAG031867
NIHAG042095
UCLA Jonsson Comprehensive Cancer CenterUNSPECIFIED
National Cancer InstituteUNSPECIFIED
Subject Keywords:AAA ATPase; p97/VCP; MSP1; p47; steady-state kinetics
Issue or Number:14
PubMed Central ID:PMC4394316
Record Number:CaltechAUTHORS:20150318-093025125
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20150318-093025125
Official Citation:Xiaoyi Zhang, Lin Gui, Xiaoyan Zhang, Stacie L. Bulfer, Valentina Sanghez, Daniel E. Wong, YouJin Lee, Lynn Lehmann, James Siho Lee, Pei-Yin Shih, Henry J. Lin, Michelina Iacovino, Conrad C. Weihl, Michelle R. Arkin, Yanzhuang Wang, and Tsui-Fen Chou Altered cofactor regulation with disease-associated p97/VCP mutations PNAS 2015 112 (14) E1705-E1714; published ahead of print March 16, 2015, doi:10.1073/pnas.1418820112
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:55882
Collection:CaltechAUTHORS
Deposited By: George Porter
Deposited On:18 Mar 2015 17:12
Last Modified:03 Oct 2019 08:09

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