We were able to identify the presence of the repeat in seven A-supergroup Wolbachia genomes (wHa, wRi, wWil, wAna, wUni, wSuzi and wGmm; see Table 1), albeit in variable copy numbers. In the Drosophila associated Wolbachia strains, the copy numbers were around 20 per genome (Table 1), whereas the other two A-supergroup genomes (wUni and wGmm) contained about half #Selleck GANT61 randurls[1|1|,|CHEM1|]# the amount of copies. Low number of hits in wUni is most likely explained by the incomplete status of the genome resulting in an underestimation of the actual copy number. In the B- (wNo, wVitB, wPip), C- (wOo, wOv), and D-supergroup (wBm) genomes, ARM was not found. Even though some

of the genomes in supergroups B, C, and D are incomplete, the total absence of the repeat in all genomes from these supergroups suggests that this motif might be Wolbachia A-supergroup

specific. Additionally, VNTR-tandem repeats associated with ARM in A-supergroup infections are also absent from genomes of B- to D-supergroups, further indicating that this feature might indeed be A-supergroup specific. Figure 1 Schematic presentation of ARM. (A) Position of ARM in association with VNTR-105 locus plus flanking regions in the wMel genome (GenBank NC_002978). Scheme for VNTR-105 repeat region was adapted from [13] (see this mTOR signaling pathway publication for detailed description of VNTR-105 structural features). Black arrows indicate the full 105 bp core repeat segment. Dashed box represents a disrupted segment. ARM (highlighted in yellow) is located within the intergenic Telomerase region containing the VNTR-105 repeat region. ARM plus repeat region are flanked by WD_1129 (red; NADH-ubiquinone oxidoreductase, putative) on the 5’-prime end and WD_1131 (green; conserved hypothetical protein, degenerate) on the 3’-prime end. (B) Detailed scheme of ARM. The 315 bp PCR amplicon is generated by primer ARM-F (21-mer) and ARM-R (18-mer). Both primers are

displayed above and below the PCR amplicon (indicated in yellow). Table 1 Number of matches to ARM in complete and draft Wolbachia genomes Wolbachia Supergroup Host Number of matches to ARM GenBank references w Mel A Drosophila melanogaster 24 NC_002978; [8] w Ha A Drosophila simulans 23 CP003884; [23] w Ri A Drosophila simulans 21 NC_012416; [22] w Wil A Drosophila willistoni 17a ASM15358v1; TSC#14030-0811.24 w Ana A Drosophila ananassae 20a ASM16747v1; [24] w Uni A Muscidifax uniraptor 7a wUni_1.0; [22] w Suzi A Drosophila suzukii 23a CAOU02000000; [25] w Gmm A Glossina morsitans morsitans 20a [14] w No B Drosophila simulans 0b CP003883; [23] w VitB B Nasonia vitripennis 0b WVB_1.0; [26] w Pip B Culex quinquefasciatus 0b NC_010981.1; [27] w Oo C Onchocerca ochengi 0b NC_018267.