Overlapping and Specific Functions of the Hsp104 N Domain Define Its Role in Protein Disaggregation
Abstract
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:
-
- Baylor College of Medicine, Houston, TX (United States). Dept. of Biochemistry and Molecular Biology
- Baylor College of Medicine, Houston, TX (United States). Dept. of Molecular and Cellular Biology
- Baylor College of Medicine, Houston, TX (United States). Dept. of Structural and Computational Biology
- Argonne National Lab. (ANL), Argonne, IL (United States). Structural Biology Center
- 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:
- Research Org.:
- Argonne National Laboratory (ANL), Argonne, IL (United States)
- Sponsoring Org.:
- USDOE Office of Science (SC), Biological and Environmental Research (BER); National Institutes of Health (NIH); Welch Foundation
- OSTI Identifier:
- 1393583
- Grant/Contract Number:
- AC02-06CH11357
- Resource 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
- Country of Publication:
- United States
- Language:
- English
- Subject:
- 59 BASIC BIOLOGICAL SCIENCES
Citation Formats
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., 2017.
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. https://doi.org/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. Mon .
"Overlapping and Specific Functions of the Hsp104 N Domain Define Its Role in Protein Disaggregation". United States. https://doi.org/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 = {Mon Sep 11 00:00:00 EDT 2017},
month = {Mon Sep 11 00:00:00 EDT 2017}
}
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