Allelic Variation at tRNA Genes in Three Nematode Species Indicates Mutation Load Despite Strong Purifying Selection

Cytosolic transfer RNAs (tRNAs), which are encoded as hundreds of genes in nuclear genomes, experience exceptionally high mutation rates and have been hypothesized to confer substantial mutational load in natural populations. Although this phenomenon appears universal across multicellular eukaryotes, a comprehensive characterization of standing variation in tRNA repertoires is still lacking in any system. Here, we resolve within-species allelic variation in nuclear-encoded tRNAs in three nematode species: Caenorhabditis elegans, C. briggsae, and C. tropicalis. We show that these genes carry signatures of high rates of historical transcription-associated mutagenesis and of purifying selection, resulting in allelic variation that includes pervasive instances of within-gene mismatches between the amino acid recognized by the tRNA backbone and that indicated by the anticodon. Furthermore, patterns of tRNA genomic organization and variation differ markedly from those of protein-coding regions. Individual genomes harbor distinct complements of tRNA genes with predicted functional differences, an observation that coincides with recent evidence that variation in tRNA expression and regulation contributes to human disease. Our findings offer an entry point for identifying the microevolutionary processes that act on tRNA repertoires and, in turn, connecting those processes to the macroevolutionary patterns that have more frequently been the focus of study.