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Cited by 3Heat shock proteins researchProtein Structure and DynamicsPhysiological and biochemical adaptations

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Pervasive Divergence in Protein Thermostability is Mediated by Both Structural Changes and Cellular Environments

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Walunjkar, Nilima, Lai, Timothy Y, Akhter, Nasima, Miller, James H, Bettinger, John Q, Marcus, Erin, Phizicky, Eric M, Ghaemmaghami, Sina, Fay, Justin C·2025·Molecular Biology and Evolution
DOI 10.1093/molbev/msaf137PMC PMC12227239

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Abstract

Temperature is a universal environmental constraint and organisms have evolved diverse mechanisms of thermotolerance. A central feature of thermophiles relative to mesophiles is a universal shift in protein stability, implying that it is a major constituent of thermotolerance. However, organisms have also evolved extensive buffering systems, such as those that disaggregate and refold denatured proteins and enable survival of heat shock. Here, we show that both cellular and protein structural changes contribute to divergence in protein thermostability between two closely related Saccharomyces species that differ by 8 °C in their thermotolerance. Using thermal proteomic profiling we find that 85% of S. cerevisiae proteins are more stable than their S. uvarum homologs and there is a 1.6 °C shift in average protein melting temperature. In an interspecific hybrid of the two species, S. cerevisiae proteins retain their thermostability, while the thermostability of their S. uvarum homologs is enhanced, indicating that cellular context contributes to protein stability differences. By purifying orthologous proteins, we show that amino acid substitutions underlie melting temperature differences for two proteins, Guk1 and Aha1. Amino acid substitutions are also computationally predicted to contribute to stability differences for most of the proteome. Our results imply that widespread changes in protein thermostability accompany the evolution of thermotolerance between closely related species.

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Metadata

Authors
Walunjkar, N., Lai, T. Y., Akhter, N., Miller, J. H., Bettinger, J. Q., Marcus, E., Phizicky, E. M., Ghaemmaghami, S., & Fay, J. C.
Affiliations
  • University of Rochester (US)
  • University of Rochester Medicine (US)
Corresponding author
Fay, Justin C
ORCID
Venue
Molecular Biology and Evolution
Publisher
Oxford University Press (OUP)
Published
2025-06-06
Volume
42
Issue
7
ISSN
  • 0737-4038 (ISSN-L)
  • 1537-1719
DOI
10.1093/molbev/msaf137
PMC
PMC12227239
OpenAlex
W4411095161
Cited by
3
References
94
Citations by year
2026: 1 · 2025: 2
FWCI
0.90
Type
journal article
Language
en
Open access
Licence
Locations
  • Molecular Biology and Evolution (publishedVersion, cc-by)
  • bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
Funders
National Institutes of Health (GM080669, GM052347, GM119502, OD025242)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Heat shock proteins research
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Structure and Dynamics
  • Physical Sciences › Environmental Science › Ecology › Physiological and biochemical adaptations
Keywords
protein thermal stability, heat tolerance, Saccharomyces cerevisiae, Saccharomyces uvarum, interspecific hybrids, thermal proteome profiling, protein melting temperature, amino acid substitutions, Aha1, cellular context, protein refolding, heat shock, thermophilic adaptation
MeSH terms
Thermotolerance, Fungal Proteins, Saccharomyces, Saccharomyces cerevisiae, Evolution, Molecular, Amino Acid Substitution, Saccharomyces cerevisiae Proteins, Proteomics, Protein Stability

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