Coevolution between Stop Codon Usage and Release Factors in Bacterial Species

Three stop codons in bacteria represent different translation termination signals, and their usage is expected to depend on their differences in translation termination efficiency, mutation bias, and relative abundance of release factors (RF1 decoding UAA and UAG, and RF2 decoding UAA and UGA). In 14 bacterial species (covering Proteobacteria, Firmicutes, Cyanobacteria, Actinobacteria and Spirochetes) with cellular RF1 and RF2 quantified, UAA is consistently over-represented in highly expressed genes (HEGs) relative to lowly expressed genes (LEGs), whereas UGA usage is the opposite even in species where RF2 is far more abundant than RF1. UGA usage relative to UAG increases significantly with P(RF2) [=RF2/(RF1 + RF2)] as expected from adaptation between stop codons and their decoders. P(RF2) is > 0.5 over a wide range of AT content (measured by P(AT3) as the proportion of AT at third codon sites), but decreases rapidly toward zero at the high range of P(AT3). This explains why bacterial lineages with high P(AT3) often have UGA reassigned because of low RF2. There is no indication that UAG is a minor stop codon in bacteria as claimed in a recent publication. The claim is invalid because of the failure to apply the two key criteria in identifying a minor codon: (1) it is least preferred by HEGs (or most preferred by LEGs) and (2) it corresponds to the least abundant decoder. Our results suggest a more plausible explanation for why UAA usage increases, and UGA usage decreases, with P(AT3), but UAG usage remains low over the entire P(AT3) range.