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bioRxiv · 10.1101/2025.09.02.673881

Modeling the Relationship between the Capsid Spike Protein Stability and Fitness in {varphi}X174 Bacteriophage

Sapozhnikov, Y.·Van Leuven, J. T.·Patel, J. S.·Miller, C. R.
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Abstract

BackgroundProtein function depends on the stable folding or binding of peptide chains, and the extent to which an amino acid substitution disrupts this stability can be a strong predictor of the mutations impact on the proteins function and the organisms fitness. This study seeks to understand this relationship in bacteriophage {phi}X174 using an experimental technique of deep mutational scanning and computationally predicted protein stability. ResultsAnalyzing the viability data using a newly developed multistage binomial model confirms that highly destabilizing mutations predict inviability. For single-site variants, the model predicts that {Delta}{Delta}G of folding greater than ~3 kcal/mol and {Delta}{Delta}G of binding greater than ~6 kcal/mol cuts the probability of survival by half of the maximum. The maximum probability, even for minimal {Delta}{Delta}G values, plateaus at around 87%, reflecting the influence of unobserved factors. Fitting of double-site variants data shows similar results. In contrast, a linear regression analysis of viable mutants reveals that the effect of protein stability is too weak to be a reliable predictor of fitness. ConclusionIn this study, we introduce a novel modeling technique to explore the relationship between protein stability and survival outcomes. While large destabilization often indicates inviability, most destabilizing mutations exert only small to moderate effects and do not reliably predict survival. Although protein stability is a crucial biophysical property influencing biological systems, it represents just one of multiple factors that contribute to fitness and survival.

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Sapozhnikov, Y., Van Leuven, J. T., Patel, J. S., Miller, C. R.. 2025-09-07. Modeling the Relationship between the Capsid Spike Protein Stability and Fitness in {varphi}X174 Bacteriophage. https://doi.org/10.1101/2025.09.02.673881

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