Abstract
Efforts to determine the mosquito genes that affect dengue virus replication have identified a number of candidates that positively or negatively modify amplification in the invertebrate host. We used deep sequencing to compare the differential transcript abundances in Aedes aegypti 14 days post dengue infection to those of uninfected A. aegypti. The gene lethal(2)-essential-for-life [l(2)efl], which encodes a member of the heat shock 20 protein (HSP20) family, was upregulated following dengue virus type 2 (DENV-2) infection in vivo. The transcripts of this gene did not exhibit differential accumulation in mosquitoes exposed to insecticides or pollutants. The induction and overexpression of l(2)efl gene products using poly(I:C) resulted in decreased DENV-2 replication in the cell line. In contrast, the RNAi-mediated suppression of l(2)efl gene products resulted in enhanced DENV-2 replication, but this enhancement occurred only if multiple l(2)efl genes were suppressed. l(2)efl homologs induce the phosphorylation of eukaryotic initiation factor 2α (eIF2α) in the fruit fly Drosophila melanogaster, and we confirmed this finding in the cell line. However, the mechanism by which l(2)efl phosphorylates eIF2α remains unclear. We conclude that l(2)efl encodes a potential anti-dengue protein in the vector mosquito.
Highlights
Aedes aegypti is an important vector of human disease pathogens, and it transmits dengue viruses that cause a spectrum of diseases that are of increasing concern as emerging infections [1,2]
We identified a family of genes, including lethal(2)-essential-for-life [l(2)efl], which encodes a product belonging to the heat shock protein 20 (HSP20) family and whose transcripts are more abundant in infected mosquitoes than in uninfected controls
A total of 10 genes identified by RNA-seq to be accumulated differentially between uninfected and infected mosquitoes were selected for real-time quantitative reverse transcriptase polymerase chain reaction (RT-PCR) analysis
Summary
Aedes aegypti is an important vector of human disease pathogens, and it transmits dengue viruses that cause a spectrum of diseases that are of increasing concern as emerging infections [1,2]. The mosquito transcriptome profiles in response to dengue infection have been studied using microarrays in cells [16], dissected organs [17,18], or whole-body preparations [11]. Specific genes, such as those related to immunity or innate immune pathways have been investigated [19,20]. The induction or inhibition of the gene and family members in vitro results in decreased and increased DENV-2 replication, respectively
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