UMass Chan Medical School Faculty Publications

UMMS Affiliation

Department of Biochemistry and Molecular Pharmacology

Publication Date

2021-08-25

Document Type

Article Preprint

Disciplines

Amino Acids, Peptides, and Proteins | Fungi | Molecular Biology

Abstract

Hsp70 interactions are critical for cellular viability and the response to stress. Previous attempts to characterize Hsp70 interactions have been limited by their transient nature and inability of current technologies to distinguish direct vs bridged interactions. We report the novel use of cross-linking mass spectrometry (XL-MS) to comprehensively characterize the budding yeast Hsp70 protein interactome. Using this approach, we have gained fundamental new insights into Hsp70 function, including definitive evidence of Hsp70 self-association as well as multi-point interaction with its client proteins. In addition to identifying a novel set of direct Hsp70 interactors which can be used to probe chaperone function in cells, we have also identified a suite of PTM-associated Hsp70 interactions. The majority of these PTMs have not been previously reported and appear to be critical in the regulation of client protein function. These data indicate that one of the mechanisms by which PTMs contribute to protein function is by facilitating interaction with chaperones. Taken together, we propose that XL-MS analysis of chaperone complexes may be used as a unique way to identify biologically-important PTMs on client proteins.

  • In vivo confirmation of Hsp70 dimerization

  • Comprehensive direct interactome of Hsp70

  • Multi-domain interactions between Hsp70 and client proteins

  • Identification of novel biologically-important client protein PTMs

Keywords

Hsp70, Ssa1, chaperone, cross-linking, mass spectrometry, posttranslational modifications, interactions, proteomics

Rights and Permissions

The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.

DOI of Published Version

10.1101/2021.08.25.457671

Source

bioRxiv 2021.08.25.457671; doi: https://doi.org/10.1101/2021.08.25.457671. Link to preprint on bioRxiv.

Comments

This article is a preprint. Preprints are preliminary reports of work that have not been certified by peer review.

Full author list omitted for brevity. For the full list of authors, see article.

The PDF available for download is Version 2 of this preprint. The complete version history of this preprint is available at https://doi.org/10.1101/2021.08.25.457671.

Journal/Book/Conference Title

bioRxiv

Creative Commons License

Creative Commons Attribution-Noncommercial-No Derivative Works 4.0 License
This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 4.0 License.

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