Temperature Adaptation at Homologous Sites in Proteins from Nine Thermophile–Mesophile Species Pairs

Whether particular amino acids are favored by selection at high temperatures over others has long been an open question in protein evolution. One way to approach this question is to compare homologous sites in proteins from one thermophile and a closely related mesophile; asymmetrical substitution patterns have been taken as evidence for selection favoring certain amino acids over others. However, most pairs of prokaryotic species that differ in optimum temperature also differ in genome-wide GC content, and amino acid content is known to be associated with GC content. Here, I compare homologous sites in nine thermophilic prokaryotes and their mesophilic relatives, all with complete published genome sequences. After adjusting for the effects of differing GC content with logistic regression, 139 of the 190 pairs of amino acids show significant substitutional asymmetry, evidence of widespread adaptive amino acid substitution. The patterns are fairly consistent across the nine pairs of species (after taking the effects of differing GC content into account), suggesting that much of the asymmetry results from adaptation to temperature. Some amino acids in some species pairs deviate from the overall pattern in ways indicating that adaptation to other environmental or physiological differences between the species may also play a role. The property that is best correlated with the patterns of substitutional asymmetry is transfer free energy, a measure of hydrophobicity, with more hydrophobic amino acids favored at higher temperatures. The correlation of asymmetry and hydrophobicity is fairly weak, suggesting that other properties may also be important.