Australian Infectious Disease Research Centre, School of Biological Sciences, The University of Queensland, Brisbane, Queensland, Australia.
Commun Biol. 2022 Jun 20;5(1):607. doi: 10.1038/s42003-022-03566-8.
The N-methyladenosine (mA) modification of RNA has been reported to affect viral infections. Studies have confirmed the role of mA in replication of several vector-borne flaviviruses, including dengue virus (DENV), in mammalian cells. Here, we explored the role of mA in DENV replication in the mosquito Aedes aegypti Aag2 cell line. We first determined the presence of mA on the RNAs from mosquito cells and using methylated RNA immunoprecipitation and sequencing (MeRIP-Seq) identified mA modification of the mosquito transcriptome and those that changed upon DENV infection. Depletion of mA methyltransferases and the mA binding protein YTHDF3 RNAs decreased the replication of DENV. In particular, we found that the Ae. aegypti ubiquitin carrier protein 9 (Ubc9) is mA 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 mA modification, implying that mA modification of Ubc9 might enhance expression of the gene. We also show that the genome of DENV is mA modified at five sites in mosquito cells. Altogether, this work reveals the involvement of mA modification in Ae. aegypti-DENV interaction.
RNA 的 N6-甲基腺苷（mA）修饰已被报道影响病毒感染。研究证实了 mA 在几种虫媒黄病毒（包括登革热病毒（DENV））在哺乳动物细胞中的复制中的作用。在这里，我们探讨了 mA 在蚊子 Aedes aegypti Aag2 细胞系中 DENV 复制中的作用。我们首先确定了蚊子细胞中 RNA 上 mA 的存在，并使用甲基化 RNA 免疫沉淀和测序（MeRIP-Seq）鉴定了蚊子转录组中的 mA 修饰以及感染 DENV 后发生变化的修饰。mA 甲基转移酶和 mA 结合蛋白 YTHDF3 RNA 的耗竭降低了 DENV 的复制。特别是，我们发现埃及伊蚊泛素载体蛋白 9（Ubc9）被 mA 修饰，并且在 DENV 感染后其表达增加。该基因的沉默和 Ubc9 的异位表达分别导致 DENV 复制减少和增加。mA 修饰抑制后 Ubc9 mRNA 的丰度及其稳定性降低，暗示 Ubc9 的 mA 修饰可能增强该基因的表达。我们还表明，DENV 的基因组在蚊子细胞中被 mA 修饰了五个位点。总之，这项工作揭示了 mA 修饰参与了埃及伊蚊-DENV 的相互作用。
