Mutations in the Yeast Hsp70, Ssa1, at P417 Alter ATP Cycling, Interdomain Coupling, and Specific Chaperone Functions

作者: Patrick G. Needham , Hardik J. Patel , Gabriela Chiosis , Patrick H. Thibodeau , Jeffrey L. Brodsky

DOI: 10.1016/J.JMB.2015.04.010

关键词:

摘要: The major cytoplasmic Hsp70 chaperones in the yeast Saccharomyces cerevisiae are Ssa proteins, and much of our understanding biology has emerged from studying ssa mutant strains. For example, Ssa1 catalyzes multiple cellular functions, including protein transport degradation, to this end, ssa1-45 proved invaluable. However, biochemical defects associated with corresponding Ssa1-45 (P417L) unknown. Consequently, we characterized P417L, as well a P417S variant, which corresponds mutation gene encoding mitochondrial Hsp70. We discovered that P417L proteins exhibit accelerated ATPase activity was similar Hsp40-stimulated rate ATP hydrolysis wild-type Ssa1. also found were compromised for peptide binding. These data consistent peptide-stimulated results limited proteolysis experiments, indicated mutants' substrate binding domains highly vulnerable digestion. Defects reactivation heat-denatured luciferase evident. Correspondingly, expressing or only copy temperature sensitive exhibited Ssa1-dependent translocation misfolded degradation. Together, studies suggest structure domain is altered coupling between nucleotide disabled when conserved P417 residue mutated. Our provide new insights into nature many allele.

参考文章(99)
Jeffrey Brodsky, Gabriela Chiosis, Hsp70 molecular chaperones: emerging roles in human disease and identification of small molecule modulators. Current Topics in Medicinal Chemistry. ,vol. 6, pp. 1215- 1225 ,(2006) , 10.2174/156802606777811997
Jianwen Jiang, Kondury Prasad, Eileen M. Lafer, Rui Sousa, Structural Basis of Interdomain Communication in the Hsc70 Chaperone Molecular Cell. ,vol. 20, pp. 513- 524 ,(2005) , 10.1016/J.MOLCEL.2005.09.028
William E. Balch, Richard I. Morimoto, Andrew Dillin, Jeffery W. Kelly, Adapting proteostasis for disease intervention. Science. ,vol. 319, pp. 916- 919 ,(2008) , 10.1126/SCIENCE.1141448
X. Zhu, X. Zhao, W. F. Burkholder, A. Gragerov, C. M. Ogata, M. E. Gottesman, W. A. Hendrickson, Structural Analysis of Substrate Binding by the Molecular Chaperone DnaK Science. ,vol. 272, pp. 1606- 1614 ,(1996) , 10.1126/SCIENCE.272.5268.1606
W. F. Burkholder, X. Zhao, X. Zhu, W. A. Hendrickson, A. Gragerov, M. E. Gottesman, Mutations in the C-terminal fragment of DnaK affecting peptide binding Proceedings of the National Academy of Sciences of the United States of America. ,vol. 93, pp. 10632- 10637 ,(1996) , 10.1073/PNAS.93.20.10632
Avrom J Caplan, Douglas M Cyr, Michael G Douglas, None, YDJ1p facilitates polypeptide translocation across different intracellular membranes by a conserved mechanism. Cell. ,vol. 71, pp. 1143- 1155 ,(1992) , 10.1016/S0092-8674(05)80063-7
Bernd Bukau, Jonathan Weissman, Arthur Horwich, Molecular Chaperones and Protein Quality Control Cell. ,vol. 125, pp. 443- 451 ,(2006) , 10.1016/J.CELL.2006.04.014
Rainer Schlecht, Sebastian R. Scholz, Heike Dahmen, Ansgar Wegener, Christian Sirrenberg, Djordje Musil, Joerg Bomke, Hans-Michael Eggenweiler, Matthias P. Mayer, Bernd Bukau, Functional Analysis of Hsp70 Inhibitors PLoS ONE. ,vol. 8, pp. e78443- ,(2013) , 10.1371/JOURNAL.PONE.0078443
A. Erbse, M.P. Mayer, B. Bukau, Mechanism of substrate recognition by Hsp70 chaperones. Biochemical Society Transactions. ,vol. 32, pp. 617- 621 ,(2004) , 10.1042/BST0320617
Ezra V. Pierpaoli, Serge M. Gisler, Philipp Christen, Sequence-specific rates of interaction of target peptides with the molecular chaperones DnaK and DnaJ. Biochemistry. ,vol. 37, pp. 16741- 16748 ,(1998) , 10.1021/BI981762Y