Ryu, Seung W. and Stewart, Rose and Pectol, D. Chase and Ender, Nicolette A. and Wimalarathne, Oshadi and Lee, Ji-Hoon and Zanini, Carlos P. and Harvey, Antony and Huibregtse, Jon M. and Mueller, Peter and Paull, Tanya T. and Walters, Kylie J. (2020) Proteome-wide identification of HSP70/HSC70 chaperone clients in human cells. PLOS Biology, 18 (7). e3000606. ISSN 1545-7885
file_id=10.1371%2Fjournal.pbio.3000606&type=printable - Published Version
Download (2MB)
Abstract
The 70 kDa heat shock protein (HSP70) family of chaperones are the front line of protection from stress-induced misfolding and aggregation of polypeptides in most organisms and are responsible for promoting the stability, folding, and degradation of clients to maintain cellular protein homeostasis. Here, we demonstrate quantitative identification of HSP70 and 71 kDa heat shock cognate (HSC70) clients using a ubiquitin-mediated proximity tagging strategy and show that, despite their high degree of similarity, these enzymes have largely nonoverlapping specificities. Both proteins show a preference for association with newly synthesized polypeptides, but each responds differently to changes in the stoichiometry of proteins in obligate multi-subunit complexes. In addition, expression of an amyotrophic lateral sclerosis (ALS)-associated superoxide dismutase 1 (SOD1) mutant protein induces changes in HSP70 and HSC70 client association and aggregation toward polypeptides with predicted disorder, indicating that there are global effects from a single misfolded protein that extend to many clients within chaperone networks. Together these findings show that the ubiquitin-activated interaction trap (UBAIT) fusion system can efficiently isolate the complex interactome of HSP chaperone family proteins under normal and stress conditions.
Item Type: | Article |
---|---|
Subjects: | Eurolib Press > Biological Science |
Depositing User: | Managing Editor |
Date Deposited: | 12 Jan 2023 07:09 |
Last Modified: | 15 Sep 2023 05:03 |
URI: | http://info.submit4journal.com/id/eprint/1196 |