Wolbachia-Induced aae-miR-12 miRNA Negatively Regulates the Expression of MCT1 and MCM6 Genes in Wolbachia-Infected Mosquito Cell Line

Abstract

BackgroundBest recognized for its role in manipulating host reproduction, the parasitic gram-negative Wolbachia pipientis is known to colonize a wide range of invertebrates. The endosymbiotic bacterium has recently been shown to cause a life-shortening effect as well as inhibiting replication of arboviruses in Aades aegypti; although the molecular mechanisms behind these effects are largely unknown. MicroRNAs (miRNAs) have been determined to have a wide range of roles in regulating gene expression in eukaryotes. A recent study showed that several A. aegypti mosquito miRNAs are differentially expressed when infected with Wolbachia. Methodology/Principal FindingsBased on the prior knowledge that one of these miRNAs, aae-miR-12, is differentially expressed in mosquitoes infected with Wolbachia, we aimed to determine any significance of this mediation. We also set out to characterize the target genes of this miRNA in the A. aegpyti genome. Bioinformatic approaches predicted a list of potential target genes and subsequent functional analyses confirmed that two of these, DNA replication licensing (MCM6) and monocarboxylate transporter (MCT1), are under the regulative control of aae-miR-12. We also demonstrated that aae-miR-12 is critical in the persistence of Wolbachia in the host cell.Conclusions/SignificanceOur study has identified two target genes of aae-miR-12, a differentially expressed mosquito miRNA in Wolbachia-infected cells, and determined that the miRNA affects Wolbachia density in the host cells.

Wolbachia: Can we save lives with a great pandemic?

“…These conflicting results will hopefully be resolved in coming years by comparative studies of species that do not receive pathogen protection from Wolbachia infection [43,67] or are actually weakened by the bacteria [68]. Finally, recent evidence suggests that Wolbachia- associated expression of host miRNAs [69,70] assists regulation of dengue virus titers within mosquitoes [71]. …”

Mosquito-specific microRNA-1174 targets serine hydroxymethyltransferase to control key functions in the gut

“…Lineage-specific miRNAs may contribute to developmental novelties in blood-feeding mosquito species and represent attractive targets for vector control approaches. Although several mosquito miRNAs have been characterized functionally (10)(11)(12)(13)(14), little is known regarding the role of mosquito-specific miRNAs and their contribution to mosquitospecific events (5).…”

Small RNAs: a new frontier in mosquito biology

“…In the same experiment, aae-miR-12 was upregulated in Ae. aegypti infected with w MelPop-CLA [23]. Two candidate target genes, DNA replication licensing factor MCM6 and monocarboxylate transporter MCT1, were bioinformatically and experimentally identified to contain aae-miR-12 complementary sites within their open reading frames (ORF).…”