Impact of translational error-induced and error-free misfolding on the rate of protein evolution

What determines the rate of protein evolution is a fundamental question in biology. Recent genomic studies revealed a surprisingly strong anticorrelation between the expression level of a protein and its rate of sequence evolution. This observation is currently explained by the translational robustness hypothesis in which the toxicity of translational error‐induced protein misfolding selects for higher translational robustness of more abundant proteins, which constrains sequence evolution. However, the impact of error‐free protein misfolding has not been evaluated. We estimate that a non‐negligible fraction of misfolded proteins are error free and demonstrate by a molecular‐level evolutionary simulation that selection against protein misfolding results in a greater reduction of error‐free misfolding than error‐induced misfolding. Thus, an overarching protein‐misfolding‐avoidance hypothesis that includes both sources of misfolding is superior to the translational robustness hypothesis. We show that misfolding‐minimizing amino acids are preferentially used in highly abundant yeast proteins and that these residues are evolutionarily more conserved than other residues of the same proteins. These findings provide unambiguous support to the role of protein‐misfolding‐avoidance in determining the rate of protein sequence evolution.