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Title: Roles of intramolecular and intermolecular interactions in functional regulation of the Hsp70 J-protein co-chaperone sis1

Journal Article · · Journal of Molecular Biology
 [1];  [1];  [2];  [1];  [3];  [2];  [4];  [1]
  1. Univ. of Wisconsin, Madison, WI (United States)
  2. Argonne National Lab. (ANL), Argonne, IL (United States)
  3. Univ. of Gdansk and Medical Univ. of Gdansk, Gdansk (Poland)
  4. Argonne National Lab. (ANL), Argonne, IL (United States); Univ. of Chicago, Chicago, IL (United States)
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Unlike other Hsp70 molecular chaperones, those of the eukaryotic cytosol have four residues, EEVD, at their C-termini. EEVD(Hsp70) binds adaptor proteins of the Hsp90 chaperone system and mitochondrial membrane preprotein receptors, thereby facilitating processing of Hsp70-bound clients through protein folding and translocation pathways. Among J-protein co-chaperones functioning in these pathways Sis1 is unique, as it also binds the EEVD(Hsp70) motif. However, little is known about the role of the Sis1:EEVD(Hsp70) interaction. We found that deletion of EEVD(Hsp70) abolished the ability of Sis1, but not the ubiquitous J-protein Ydj1, to partner with Hsp70 in in vitro protein refolding. Sis1 co-chaperone activity with Hsp70ΔEEVD was restored upon substitution of a glutamic acid of the J-domain. Structural analysis revealed that this key glutamic acid, which is not present in Ydj1, forms a salt bridge with an arginine of the immediately adjacent glycine-rich region. Thus, restoration of Sis1 in vitro activity suggests that intramolecular interaction(s) between the J-domain and glycine-rich region controls co-chaperone activity, which is optimal only when Sis1 interacts with the EEVD(Hsp70) motif. Yet, we found that disruption of the Sis1:EEVD(Hsp70) interaction enhances the ability of Sis1 to substitute for Ydj1 in vivo. Our results are consistent with the idea that interaction of Sis1 with EEVD(Hsp70) minimizes transfer of Sis1-bound clients to Hsp70s that are primed for client transfer to folding and translocation pathways by their preassociation with EEVD-binding adaptor proteins. Finally, these interactions may be one means by which cells triage Ydj1- and Sis1-bound clients to productive and quality control pathways, respectively.

Research Organization:
Argonne National Laboratory (ANL), Argonne, IL (United States)
Sponsoring Organization:
USDOE Office of Science (SC), Biological and Environmental Research (BER)
Grant/Contract Number:
AC02-06CH11357; GM31107; GM094585; 2013/09/N/NZ2/01979
OSTI ID:
1263634
Alternate ID(s):
OSTI ID: 1367767
Journal Information:
Journal of Molecular Biology, Vol. 427, Issue 7; ISSN 0022-2836
Publisher:
ElsevierCopyright Statement
Country of Publication:
United States
Language:
English
Citation Metrics:
Cited by: 31 works
Citation information provided by
Web of Science

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Cited By (13)

“Conformational lock” via unusual intramolecular C–F⋯OC and C–H⋯Cl–C parallel dipoles observed in in situ cryocrystallized liquids journal January 2016
The Hsp70 chaperone network journal June 2019
The C-terminal GGAP motif of Hsp70 mediates substrate recognition and stress response in yeast journal September 2018
Structural and functional analysis of the Hsp70/Hsp40 chaperone system journal October 2019
Graphene-based materials do not impair physiology, gene expression and growth dynamics of the aeroterrestrial microalga Trebouxia gelatinosa journal February 2019
Genome-wide identification and expression analysis of the molecular chaperone binding protein BiP genes in Citrus journal November 2018
Solution structure of the J domain of DnaJ homolog subfamily B member 8 dataset January 2010
Graphene-based materials do not impair physiology, gene expression and growth dynamics of the aeroterrestrial microalga Trebouxia gelatinosa text January 2019
Graphene-based materials do not impair physiology, gene expression and growth dynamics of the aeroterrestrial microalga Trebouxia gelatinosa text January 2019
Functionality of Class A and Class B J‐protein co‐chaperones with Hsp70 journal August 2015
The Plasmodium falciparum Hsp70‐x chaperone assists the heat stress response of the malaria parasite journal November 2019
Broadening the functionality of a J-protein/Hsp70 molecular chaperone system journal October 2017
Client processing is altered by novel myopathy-causing mutations in the HSP40 J domain journal June 2020

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Related Subjects

59 BASIC BIOLOGICAL SCIENCES
37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CHEMISTRY
molecular chaperone
protein folding
Hsp40
J domain
EEVD motif