Origin of an Alternative Genetic Code in the Extremely Small and GC–Rich Genome of a Bacterial Symbiont

Abstract

The genetic code relates nucleotide sequence to amino acid sequence and is shared across all organisms, with the rare exceptions of lineages in which one or a few codons have acquired novel assignments. Recoding of UGA from stop to tryptophan has evolved independently in certain reduced bacterial genomes, including those of the mycoplasmas and some mitochondria. Small genomes typically exhibit low guanine plus cytosine (GC) content, and this bias in base composition has been proposed to drive UGA Stop to Tryptophan (Stop→Trp) recoding. Using a combination of genome sequencing and high-throughput proteomics, we show that an α-Proteobacterial symbiont of cicadas has the unprecedented combination of an extremely small genome (144 kb), a GC–biased base composition (58.4%), and a coding reassignment of UGA Stop→Trp. Although it is not clear why this tiny genome lacks the low GC content typical of other small bacterial genomes, these observations support a role of genome reduction rather than base composition as a driver of codon reassignment. The genetic code, which relates DNA sequence to protein sequence, is nearly universal across all life. Examples of recodings do exist, but new instances are rare. Genomes that exhibit recodings typically have other extreme properties, including reduced size, reduced gene sets, and low guanine plus cytosine (GC) content. The most common recoding event, the reassignment of UGA to Tryptophan instead of Stop (Stop→Trp), was previously known from several mitochondrial and one bacterial lineage, and it was proposed to be driven by extinction of the UGA codon due to reduction in GC content. Here we present an unusual bacterial genome from a symbiont of cicadas. It exhibits the UGA Stop→Trp reassignment, but has a high GC content, showing that reduction in GC content is not a necessary condition for this recoding. This symbiont genome is also the smallest known for any cellular organism. We therefore propose gene loss during genome reduction as the common force driving this code change in bacteria and organelles. Additionally, the extremely small size of the genome further obscures the once-clear distinction between organelle and autonomous bacterial life.