The long-term objective of this project is to utilize the yeast S. cerevisiae as a model system to determine the mechanism by which the cytosolic Hsp40s Sis1 and Ydjl act with Hsp70 Ssal (Ssal) to protect cells from denaturing physiological stresses such as heat-shock. The hallmark of the heat-shock response is the rapid induction of chaperone protein expression and the global suppression of protein synthesis. These events occur to prevent the accumulation of non-native proteins and suppress protein aggregation. Hsp40s function to suppress protein aggregation by binding non-native proteins and delivering them to the polypeptide binding site of HspT0.
In aim 1, we propose a series of experiments that are designed to determine the mechanism for substrate binding by Hsp40s.
In aim 2, we will investigate how denatured proteins that are bound to Hsp40 are delivered to Hsp70 to facilitate protein refolding. An additional feature of Hsp40 family members is that they have evolved to have different domain structures and can target Hsp70 to catalyze different cellular reactions. We have demonstrated that the Ydjl and Sis1 function in the cytosol to direct Ssal to facilitate different aspects of protein metabolism.
In Aim 3 and Aim 4 we propose a series of domain swap experiments between Ydjl and Sis1 that are designed to define the rules by which Hsp70s cellular functions are specified.The outcome of these studies will define basic mechanisms by which Hsp40s function with Hsp70 to protect cells from physiological stress.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM067785-02
Application #
6744186
Study Section
Physiological Chemistry Study Section (PC)
Program Officer
Wehrle, Janna P
Project Start
2003-05-01
Project End
2007-04-30
Budget Start
2004-05-01
Budget End
2005-04-30
Support Year
2
Fiscal Year
2004
Total Cost
$253,681
Indirect Cost
Name
University of North Carolina Chapel Hill
Department
Anatomy/Cell Biology
Type
Schools of Medicine
DUNS #
608195277
City
Chapel Hill
State
NC
Country
United States
Zip Code
27599
Veit, Gudio; Avramescu, Radu G; Chiang, Annette N et al. (2016) From CFTR biology toward combinatorial pharmacotherapy: expanded classification of cystic fibrosis mutations. Mol Biol Cell 27:424-33
Vermulst, Marc; Denney, Ashley S; Lang, Michael J et al. (2015) Transcription errors induce proteotoxic stress and shorten cellular lifespan. Nat Commun 6:8065
Wolfe, Katie J; Ren, Hong Yu; Trepte, Philipp et al. (2014) Polyglutamine-rich suppressors of huntingtin toxicity act upstream of Hsp70 and Sti1 in spatial quality control of amyloid-like proteins. PLoS One 9:e95914
Houck, Scott A; Ren, Hong Yu; Madden, Victoria J et al. (2014) Quality control autophagy degrades soluble ERAD-resistant conformers of the misfolded membrane protein GnRHR. Mol Cell 54:166-179
Ren, Hong Yu; Grove, Diane E; De La Rosa, Oxana et al. (2013) VX-809 corrects folding defects in cystic fibrosis transmembrane conductance regulator protein through action on membrane-spanning domain 1. Mol Biol Cell 24:3016-24
Wolfe, Katie J; Ren, Hong Yu; Trepte, Philipp et al. (2013) The Hsp70/90 cochaperone, Sti1, suppresses proteotoxicity by regulating spatial quality control of amyloid-like proteins. Mol Biol Cell 24:3588-602
Borges, JĂșlio C; Seraphim, Thiago V; Mokry, David Z et al. (2012) Identification of regions involved in substrate binding and dimer stabilization within the central domains of yeast Hsp40 Sis1. PLoS One 7:e50927
Wolfe, Katie J; Cyr, Douglas M (2011) Amyloid in neurodegenerative diseases: friend or foe? Semin Cell Dev Biol 22:476-81
Silva, Julio C; Borges, Julio C; Cyr, Douglas M et al. (2011) Central domain deletions affect the SAXS solution structure and function of yeast Hsp40 proteins Sis1 and Ydj1. BMC Struct Biol 11:40
Ren, Hong Yu; Patterson, Cam; Cyr, Douglas M et al. (2011) Reconstitution of CHIP E3 ubiquitin ligase activity. Methods Mol Biol 787:93-103

Showing the most recent 10 out of 22 publications