TY - JOUR T1 - Alpha-mannosidase-2 modulates arbovirus infection in a pathogen- and <em>Wolbachia</em>-specific manner in <em>Aedes aegypti</em> mosquitoes JF - bioRxiv DO - 10.1101/2022.03.18.484928 SP - 2022.03.18.484928 AU - Nadya Urakova AU - Renuka E. Joseph AU - Allyn Huntsinger AU - Vanessa M. Macias AU - Matthew J. Jones AU - Leah T. Sigle AU - Ming Li AU - Omar S. Akbari AU - Zhiyong Xi AU - Konstantinos Lymperopoulos AU - Richard T Sayre AU - Elisabeth A. McGraw AU - Jason L. Rasgon Y1 - 2022/01/01 UR - http://biorxiv.org/content/early/2022/03/19/2022.03.18.484928.abstract N2 - Multiple Wolbachia strains can block pathogen infection, replication, and/or transmission in Aedes aegypti mosquitoes under both laboratory and field conditions. However, Wolbachia effects on pathogens can be highly variable across systems and the factors governing this variability are not well understood. It is increasingly clear that the mosquito host is not a passive player in which Wolbachia governs pathogen transmission phenotypes; rather, the genetics of the host can significantly modulate Wolbachia-mediated pathogen blocking. Specifically, previous work linked variation in Wolbachia pathogen blocking to polymorphisms in the mosquito alpha-mannosidase 2 (αMan2) gene. Here we use CRISPR-Cas9 mutagenesis to functionally test this association. We developed αMan2 knockouts and examined effects on both Wolbachia and virus levels, using both dengue virus (DENV; Flaviviridae) and Mayaro virus (MAYV; Togaviridae). Wolbachia titers were significantly elevated in αMan2 knockout (KO) mosquitoes, but there were complex interactions with virus infection and replication. In Wolbachia-uninfected mosquitoes, the αMan2 KO mutation was associated with decreased DENV titers, but in a Wolbachia-infected background, the αMan2 KO mutation significantly modulated virus blocking. In contrast, the αMan2 KO mutation significantly increased MAYV replication in Wolbachia-uninfected mosquitoes and did not affect Wolbachia-mediated virus blocking. These results demonstrate that αMan2 modulates arbovirus infection in Ae. aegypti mosquitoes in a pathogen- and Wolbachia-specific manner, and that Wolbachia-mediated pathogen blocking is a complex phenotype dependent on the mosquito host genotype and the pathogen. These results have significant impact for the design and use of Wolbachia-based strategies to control vector-borne pathogens.Competing Interest StatementThe authors have declared no competing interest.cDNAcomplementary DNACIcytoplasmic incompatibilityDENVdengue virusDMEMDulbecco’s Modified Eagle MediumDPIdays post infectionFBSFetal bovine serumFFAfocus-forming assayFFUfocus-forming unitgDNAgenomic DNAGEGenome equivalentKOknockoutMAYVMayaro virusPBpathogen blockingPBSPhosphate-buffered salinePFUPlaque-forming unitqPCRQuantitative polymerase chain reaction ER -