N6-methyladenosine modification of the Aedes aegypti transcriptome and its alteration upon dengue virus infection in Aag2 cell line

Abstract

The N6-methyladenosine (m6A) modification of RNA has been reported to affect viral infections. Studies have confirmed the role of m6A in replication of several vector-borne flaviviruses, including dengue virus (DENV), in mammalian cells. Here, we explored the role of m6A in DENV replication in the mosquito Aedes aegypti Aag2 cell line. We first determined the presence of m6A on the RNAs from mosquito cells and using methylated RNA immunoprecipitation and sequencing (MeRIP-Seq) identified m6A modification of the mosquito transcriptome and those that changed upon DENV infection. Depletion of m6A methyltransferases and the m6A binding protein YTHDF3 RNAs decreased the replication of DENV. In particular, we found that the Ae. aegypti ubiquitin carrier protein 9 (Ubc9) is m6A modified and its expression increases after DENV infection. Silencing of the gene and ectopic expression of Ubc9 led to reduced and increased DENV replication, respectively. The abundance of Ubc9 mRNA and its stability were reduced with the inhibition of m6A modification, implying that m6A modification of Ubc9 might enhance expression of the gene. We also show that the genome of DENV is m6A modified at five sites in mosquito cells. Altogether, this work reveals the involvement of m6A modification in Ae. aegypti-DENV interaction.

ALKBH8 as a potential N6‐methyladenosine (m6A) eraser in insects

“…It has been reported that the conserved region of METTL3 and METTL14 is the MT‐A70 domain, and the conserved region of YTHDC and YTHDF is the YTH domain (Du et al, 2016, p. 4; Zaccara et al, 2019). These two highly conserved domains are also present in the mosquito m 6 A‐related gene (Dai et al, 2022). Phylogenetic analysis also showed that the genes of the abovementioned four enzymes in mosquitoes have high similarity with the homologous genes in other species selected (Dai et al, 2022).…”

“…These two highly conserved domains are also present in the mosquito m 6 A‐related gene (Dai et al, 2022). Phylogenetic analysis also showed that the genes of the abovementioned four enzymes in mosquitoes have high similarity with the homologous genes in other species selected (Dai et al, 2022). In particular, all of them have the same conserved domains, and the evolutionary relationship of genes corresponds with the evolutionary level of species.…”

“…In the silkworm, Bombyx mori , knocking down BmMETTL3 and BmMETTL14 in a B. mori cell line resulted in the arrest of cell cycle progression and deficiency of chromosome alignment and segregation (Zhang et al, 2019). Previous reports demonstrated m 6 A modification of transcripts in two mosquito species, Aedes aegypti and Anopheles sinensis (Dai, Etebari, & Asgari, 2022; Liu, Cao, Zhang, Wu, & Yin, 2022). The studies further indicated that m 6 A methylation is involved in viral infection and sperm tail formation in the mosquitoes, respectively.…”

N6‐methyladenosine modification of RNA controls dopamine synthesis to influence labour division in ants

“…However, m 6 A methylation positively regulates gene expression. For instance, the abundance of ubiquitin carrier protein 9 (Ubc9) mRNA and its stability were decreased by the inhibition of m 6 A modification in Aedes aegypti (Dai et al, 2022).…”

Dynamic transcriptome analyses reveal m6A regulated immune non-coding RNAs during dengue disease progression

“…The m6A modification has been identified as an important regulator in regulating expression of lncRNAs [ 18 , 19 ]. However, limited studies have investigated the m6A modification during dengue infection [ 20 , 21 ]. It is important to understand how the host immune system reacts to infection and how the virus evades immune response for developing effective antivirals [ 22 ].…”