A single synonymous nucleotide change impacts the male-killing phenotype of prophage WO genewmk

Wolbachiaare the most widespread bacterial endosymbionts in animals. Within arthropods, these maternally-transmitted bacteria can selfishly hijack host reproductive processes to increase the relative fitness of their transmitting females. One such form of reproductive parasitism called male killing, or the selective killing of infected males, is recapitulated to degrees by transgenic expression of theWO-mediated killing(wmk) gene. Here, we characterize the genotype-phenotype landscape ofwmk-induced male killing inD.melanogasterusing transgenic expression. While phylogenetically distantwmkhomologs induce no sex-ratio bias, closely-related homologs exhibit complex phenotypes spanning no death, male death, or death of all hosts. We demonstrate that alternative start codons, synonymous codons, and notably a single synonymous nucleotide inwmkcan ablate killing. These findings reveal previously unrecognized features of transgenicwmk-induced killing and establish new hypotheses for the impacts of post-transcriptional processes in male killing variation. We conclude that synonymous sequence changes are not necessarily silent in nested endosymbiotic interactions with life-or-death consequences.