, 2008) between examined Sodalis isolates, C. melbae, and C. columbae symbionts. The ompA, ompC, and rcsF loci (Fig. 2) appear to be more informative toward the phylogenetic resolution of the Sodalis-like symbiont clade. With CX-5461 nmr rcsF, sufficient phylogenetic signal was provided to enable clustering of the Glossina symbionts, with strong support, separate from the C. melbae symbiont (Fig. 2b). Interestingly, rcsF in E. coli has been shown to be involved in signaling transduction of perturbations and/or environmental cues from the cell surface (Majdalani et al., 2005). Diversification between Sodalis and C. melbae isolates may indicate functional
adaptations, such as differences in the type of signaling encountered within the host species background. The Sodalis symbionts also formed a distinct clade with the ompC phylogeny, with most mutations noted outside of the seven putative extracellular loops (Basle et al., 2006) of the different Glossina isolates. The one exception occurred in extracellular loop 4, where host interspecies diversity was observed with Sodalis isolates. Relative to the other surface encoding genes analyzed in this study, the ompA gene exhibited the greatest diversity among symbionts due to a combination of point mutations
and indels. The best-studied ompA gene variant, that of E. coli K-12, encodes a 325 amino acid polypeptide Dimethyl sulfoxide (Chen et al., 1980). The N-terminal domain forms an eight-stranded β-barrel in the outer membrane, creating four surface-exposed loops (Pautsch & Schulz, 1998), while the C-terminus is Selleck Y 27632 periplasmic (Klose et al., 1988). Amino acid variations within outer membrane proteins mainly occur in the
domains located in the extracellular regions, while interspaced residues making up the β-strands tend to be conserved. In our analyses, relative to Glossina symbionts, a total of nine nonsynonymous mutations were observed among C. melbae, C. columbae, and Sitophilus (i.e. Sitophilus oryzae primary symbiont, SOPE) symbionts occurring in loops 1–4 of the OmpA protein. Differences noted in the ompA sequence between the Glossina symbionts were localized outside of the extracellular regions, similar to our observations with ompC. In relation to ompA, the C. columbae symbiont exhibited the greatest nucleotide divergence resulting in its sister taxon placement relative to the other symbionts of interest with strong MP bootstrap support. MP, Bayesian, and NJ analyses all grouped Glossina symbionts within their own clade indicative of diversification potentially arising from host adaptation processes. The Sodalis ompA gene demonstrated a wide nucleotide variation (π) within tsetse species (Table 1), with the highest π exhibited within G. morsitans (π=0.11) and the lowest within G. brevipalpis (π=0.001).