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Title: Overlapping and Specific Functions of the Hsp104 N Domain Define Its Role in Protein Disaggregation

Journal Article · · Scientific Reports
 [1];  [1];  [2];  [3];  [4];  [5];  [5]
  1. Baylor College of Medicine, Houston, TX (United States). Dept. of Biochemistry and Molecular Biology
  2. Baylor College of Medicine, Houston, TX (United States). Dept. of Molecular and Cellular Biology
  3. Baylor College of Medicine, Houston, TX (United States). Dept. of Structural and Computational Biology
  4. Argonne National Lab. (ANL), Argonne, IL (United States). Structural Biology Center
  5. Baylor College of Medicine, Houston, TX (United States). Dept. of Biochemistry and Molecular Biology; Baylor College of Medicine, Houston, TX (United States). Center for Drug Discovery
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Hsp104 is a ring-forming protein disaggregase that rescues stress-damaged proteins from an aggregated state. To facilitate protein disaggregation, Hsp104 cooperates with Hsp70 and Hsp40 chaperones (Hsp70/40) to form a bi-chaperone system. How Hsp104 recognizes its substrates, particularly the importance of the N domain, remains poorly understood and multiple, seemingly conficting mechanisms have been proposed. Although the N domain is dispensable for protein disaggregation, it is sensitive to point mutations that abolish the function of the bacterial Hsp104 homolog in vitro, and is essential for curing yeast prions by Hsp104 overexpression in vivo. Here, we present the crystal structure of an N-terminal fragment of Saccharomyces cerevisiae Hsp104 with the N domain of one molecule bound to the C-terminal helix of the neighboring D1 domain. Consistent with mimicking substrate interaction, mutating the putative substrate-binding site in a constitutively active Hsp104 variant impairs the recovery of functional protein from aggregates. We fnd that the observed substrate-binding defect can be rescued by Hsp70/40 chaperones, providing a molecular explanation as to why the N domain is dispensable for protein disaggregation when Hsp70/40 is present, yet essential for the dissolution of Hsp104-specifc substrates, such as yeast prions, which likely depends on a direct N domain interaction.

Research Organization:
Argonne National Laboratory (ANL), Argonne, IL (United States)
Sponsoring Organization:
USDOE Office of Science (SC), Biological and Environmental Research (BER); National Institutes of Health (NIH); Welch Foundation
Grant/Contract Number:
AC02-06CH11357
OSTI ID:
1393583
Journal Information:
Scientific Reports, Vol. 7, Issue 1; ISSN 2045-2322
Publisher:
Nature Publishing GroupCopyright Statement
Country of Publication:
United States
Language:
English
Citation Metrics:
Cited by: 10 works
Citation information provided by
Web of Science

References (50)

Roles of Individual Domains and Conserved Motifs of the AAA+ Chaperone ClpB in Oligomerization, ATP Hydrolysis, and Chaperone Activity journal March 2003
Asymmetric deceleration of ClpB or Hsp104 ATPase activity unleashes protein-remodeling activity journal January 2007
A tightly regulated molecular toggle controls AAA+ disaggregase journal November 2012
The Amino-terminal Domain of ClpB Supports Binding to Strongly Aggregated Proteins journal August 2005
In vivo aspects of protein folding and quality control journal June 2016
Ratchet-like polypeptide translocation mechanism of the AAA+ disaggregase Hsp104 journal June 2017
Structure and Activity of ClpB from Escherichia coli: ROLE OF THE AMINO- AND CARBOXYL-TERMINAL DOMAINS journal September 2000
Functional analysis of conserved cis- and trans-elements in the Hsp104 protein disaggregating machine journal August 2012
Molecular chaperones: guardians of the proteome in normal and disease states journal January 2015
Hsp70 targets Hsp100 chaperones to substrates for protein disaggregation and prion fragmentation journal August 2012
Inorganic phosphate assay with malachite green: An improvement and evaluation journal December 1982
Hsp70 proteins bind Hsp100 regulatory M domains to activate AAA+ disaggregase at aggregate surfaces journal November 2012
Aggregate reactivation mediated by the Hsp100 chaperones journal April 2012
AAA+ Proteases: ATP-Fueled Machines of Protein Destruction journal July 2011
PDBsum: a web-based database of summaries and analyses of all PDB structures journal December 1997
Heat shock protein (Hsp) 70 is an activator of the Hsp104 motor journal May 2013
ClpS, a Substrate Modulator of the ClpAP Machine journal March 2002
Structure and mechanism of the hexameric MecA–ClpC molecular machine journal March 2011
Evidence for an Unfolding/Threading Mechanism for Protein Disaggregation by Saccharomyces cerevisiae Hsp104 journal May 2004
Protein rescue from aggregates by powerful molecular chaperone machines journal September 2013
Crystal Structure of the E. coli Hsp100 ClpB N-Terminal Domain journal March 2003
ClpB N-terminal domain plays a regulatory role in protein disaggregation journal November 2015
Species-specific collaboration of heat shock proteins (Hsp) 70 and 100 in thermotolerance and protein disaggregation journal April 2011
In Vivo Monitoring of the Prion Replication Cycle Reveals a Critical Role for Sis1 in Delivering Substrates to Hsp104 journal November 2008
The Structure of ClpB journal October 2003
Hsp104, Hsp70, and Hsp40 journal July 1998
Crystal Structure of the Heterodimeric Complex of the Adaptor, ClpS, with the N-domain of the AAA + Chaperone, ClpA journal September 2002
The M-Domain Controls Hsp104 Protein Remodeling Activity in an Hsp70/Hsp40-Dependent Manner journal September 2010
Spiral architecture of the Hsp104 disaggregase reveals the basis for polypeptide translocation journal August 2016
Peptide and Protein Binding in the Axial Channel of Hsp104: INSIGHTS INTO THE MECHANISM OF PROTEIN UNFOLDING journal August 2008
Orientation of the amino-terminal domain of ClpB affects the disaggregation of the protein: Orientation of the N-terminal domain of ClpB journal March 2012
Hsp104, Hsp70 and Hsp40 interplay regulates formation, growth and elimination of Sup35 prions journal October 2008
Saccharomyces cerevisiae Hsp104 Enhances the Chaperone Capacity of Human Cells and Inhibits Heat Stress-Induced Proapoptotic Signaling † journal June 2004
The Hsp104 N-Terminal Domain Enables Disaggregase Plasticity and Potentiation journal March 2015
Potentiated Hsp104 Variants Antagonize Diverse Proteotoxic Misfolding Events journal January 2014
Molecular Basis for the Unique Role of the AAA + Chaperone ClpV in Type VI Protein Secretion journal July 2011
Arginine phosphorylation marks proteins for degradation by a Clp protease journal October 2016
CryoEM structure of Hsp104 and its mechanistic implication for protein disaggregation journal April 2010
N-Terminal Domain of Yeast Hsp104 Chaperone Is Dispensable for Thermotolerance and Prion Propagation but Necessary for Curing Prions by Hsp104 Overexpression journal April 2006
Dominant Gain-of-Function Mutations in Hsp104p Reveal Crucial Roles for the Middle Region journal May 2004
Cooperation of Hsp70 and Hsp100 chaperone machines in protein disaggregation journal May 2015
The N-terminal domain of Escherichia coli ClpB enhances chaperone function journal July 2005
The N Terminus of ClpB from Thermus thermophilus Is Not Essential for the Chaperone Activity journal September 2002
The Truncated Form of the Bacterial Heat Shock Protein ClpB/HSP100 Contributes to Development of Thermotolerance in the Cyanobacterium Synechococcus sp. Strain PCC 7942 journal December 2000
Interaction of the N-terminal domain of Escherichia coli heat-shock protein ClpB and protein aggregates during chaperone activity journal December 2004
Structural basis for the disaggregase activity and regulation of Hsp104 journal November 2016
[20] Processing of X-ray diffraction data collected in oscillation mode book January 1997
Conserved Amino Acid Residues within the Amino-terminal Domain of ClpB are Essential for the Chaperone Activity journal August 2002
Substrate threading through the central pore of the Hsp104 chaperone as a common mechanism for protein disaggregation and prion propagation journal April 2008
Site-directed mutagenesis of the dual translational initiation sites of the clpB gene of Escherichia coli and characterization of its gene products. journal September 1993

Cited By (3)

Structural determinants for protein unfolding and translocation by the Hsp104 protein disaggregase journal December 2017
Cryo-EM Structures of the Hsp104 Protein Disaggregase Captured in the ATP Conformation journal January 2019
Forms and abundance of chaperone proteins influence yeast prion variant competition: Prion Variant Competition journal February 2019

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59 BASIC BIOLOGICAL SCIENCES