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

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.
Authors:
 [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
Publication Date:
Grant/Contract Number:
AC02-06CH11357
Type:
Accepted Manuscript
Journal Name:
Scientific Reports
Additional Journal Information:
Journal Volume: 7; Journal Issue: 1; Journal ID: ISSN 2045-2322
Publisher:
Nature Publishing Group
Research Org:
Argonne National Lab. (ANL), Argonne, IL (United States)
Sponsoring Org:
USDOE Office of Science (SC), Biological and Environmental Research (BER) (SC-23); National Institutes of Health (NIH); Welch Foundation
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES
OSTI Identifier:
1393583

Lee, Jungsoon, Sung, Nuri, Mercado, Jonathan M., Hryc, Corey F., Chang, Changsoo, Lee, Sukyeong, and T. F. Tsai, Francis. Overlapping and Specific Functions of the Hsp104 N Domain Define Its Role in Protein Disaggregation. United States: N. p., Web. doi:10.1038/s41598-017-11474-9.
Lee, Jungsoon, Sung, Nuri, Mercado, Jonathan M., Hryc, Corey F., Chang, Changsoo, Lee, Sukyeong, & T. F. Tsai, Francis. Overlapping and Specific Functions of the Hsp104 N Domain Define Its Role in Protein Disaggregation. United States. doi:10.1038/s41598-017-11474-9.
Lee, Jungsoon, Sung, Nuri, Mercado, Jonathan M., Hryc, Corey F., Chang, Changsoo, Lee, Sukyeong, and T. F. Tsai, Francis. 2017. "Overlapping and Specific Functions of the Hsp104 N Domain Define Its Role in Protein Disaggregation". United States. doi:10.1038/s41598-017-11474-9. https://www.osti.gov/servlets/purl/1393583.
@article{osti_1393583,
title = {Overlapping and Specific Functions of the Hsp104 N Domain Define Its Role in Protein Disaggregation},
author = {Lee, Jungsoon and Sung, Nuri and Mercado, Jonathan M. and Hryc, Corey F. and Chang, Changsoo and Lee, Sukyeong and T. F. Tsai, Francis},
abstractNote = {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.},
doi = {10.1038/s41598-017-11474-9},
journal = {Scientific Reports},
number = 1,
volume = 7,
place = {United States},
year = {2017},
month = {9}
}