Assessing Aedes aegypti candidate genes during viral infection and Wolbachia-mediated pathogen blocking

The symbiont Wolbachia limits viral replication in Aedes aegypti and significantly reduces dengue fever incidence in humans following field release. Despite plans for its widespread use, Wolbachia’s mode of action remains poorly understood. Many studies suggest that the mechanism of viral blocking is likely multifaceted, involving aspects of immunity, cellular stress, and nutritional competition. A previous study from our group used artificial selection to identify a new mosquito candidate gene related to viral blocking; alpha-mannosidase-2a (alpha-Mann-2a) with a predicted role in protein glycosylation. Protein glycosylation pathways tend to be involved in complex host-viral interactions; however, the function of alpha-mannosidases has not been described in mosquito-virus interactions. We examined the expression of alpha-Mann-2a in response to virus and Wolbachia infections and whether reduced gene expression, caused by RNA interference (RNAi), affected viral loads. We show that dengue virus (DENV) infection affects the expression of alpha-Mann-2a in a tissue and time dependent manner, whereas Wolbachia infection had no effect. In the midgut, DENV prevalence increased following knockdown of alpha-Mann-2a expression in Wolbachia-free mosquitoes, suggesting that alpha-Mann-2a interferes with infection. Expression knockdown had the same effect on the Togavirus chikungunya (CHIKV), indicating that alpha-Mann-2a may have broad antivirus effects in the midgut. Interestingly, we were unable to knockdown the expression in Wolbachia-infected mosquitoes. We also provide evidence that alpha-Mann-2a may affect the transcriptional level of another gene predicted to be involved in viral blocking and cell adhesion; cadherin87a. These data support the hypothesis that glycosylation and adhesion pathways may broadly be involved in viral infection in Ae. aegypti.