Cited by 28Advanced Proteomics Techniques and ApplicationsHeat shock proteins researchUbiquitin and proteasome pathways

Mutant thermal proteome profiling for characterization of missense protein variants and their associated phenotypes within the proteome

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Peck Justice, Sarah A., Barron, Monica P., Qi, Guihong D., Wijeratne, H. R. Sagara, Victorino, José F., Simpson, Ed R., Vilseck, Jonah Z., Wijeratne, Aruna B. +1 more·2020·Journal of Biological Chemistry

Published version of a preprint

Abstract

rofiling (mTPP). This study characterized global impacts of temperature sensitivity-inducing missense mutations in two different subunits of the 26S proteasome. The majority of alterations identified by RNA-Seq and global proteomics were similar between the mutants, which could suggest that a similar functional disruption is occurring in both missense variants. Results from mTPP, however, provide unique insights into the mechanisms that contribute to the TS phenotype in each mutant, revealing distinct changes that were not obtained using only steady-state transcriptome and proteome analyses. Computationally, multisite λ-dynamics simulations add clear support for mTPP experimental findings. This work shows that mTPP is a precise approach to measure changes in missense mutant-containing proteomes without the requirement for large amounts of starting material, specific antibodies against proteins of interest, and/or genetic manipulation of the biological system. Although experiments were performed under permissive conditions, mTPP provided insights into the underlying protein stability changes that cause dramatic cellular phenotypes observed at nonpermissive temperatures. Overall, mTPP provides unique mechanistic insights into missense mutation dysfunction and connection of genotype to phenotype in a rapid, nonbiased fashion.

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Authors
Peck Justice, S. A., Barron, M. P., Qi, G. D., Wijeratne, H. R. S., Victorino, J. F., Simpson, E. R., Vilseck, J. Z., Wijeratne, A. B., & Mosley, A. L.
Affiliations
Indiana University Indianapolis (US)
Corresponding author
  • Wijeratne, Aruna B.
  • Mosley, Amber L.
Venue
Journal of Biological Chemistry
Publisher
Elsevier BV
Published
2020-11
Volume
295
Issue
48
Pages
16219-16238
ISSN
0021-9258 (ISSN-L)
OpenAlex
W3082270748
Cited by
28
References
156
Citations by year
2025: 4 · 2024: 6 · 2023: 3 · 2022: 6 · 2021: 7 · 2020: 2
FWCI
1.72
Type
journal article
Language
en
Open access
Locations
  • Journal of Biological Chemistry (publishedVersion, cc-by)
  • bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion, cc-by-nc-nd)
  • PubMed Central (publishedVersion)
  • PubMed (publishedVersion)
  • IUScholarWorks (Indiana University) (submittedVersion, cc-by)
Funders
  • HHS | NIH | National Heart, Lung, and Blood Institute (HL007910)
  • Ralph W. and Grace M. Showalter Research Trust Fund (N/A)
  • HHS | NIH | National Center for Advancing Translational Sciences (UL1TR002529)
  • Thermo Fisher Scientific (Reagent Award)
Topics
  • Physical Sciences › Chemistry › Spectroscopy › Advanced Proteomics Techniques and Applications
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Heat shock proteins research
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Ubiquitin and proteasome pathways
Keywords
missense mutations, temperature sensitivity, 26S proteasome, proteasome subunits, protein stability, genotype-phenotype correlation
MeSH terms
RNA-Seq, Saccharomyces cerevisiae, Temperature, Mutation, Missense, Proteome, Saccharomyces cerevisiae Proteins, Proteasome Endopeptidase Complex

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