Differential viral RNA methylation contributes to pathogen blocking in Wolbachia-colonized arthropods.
Differential viral RNA methylation contributes to pathogen blocking in Wolbachia-colonized arthropods.
复制标题
DOI：
10.1371/journal.ppat.1010393
复制
发表时间：
2022-03
期刊：
影响因子：
6.7
通讯作者：
Hardy RW
中科院分区：
文献类型：
--
作者：
Bhattacharya T;Yan L;Crawford JM;Zaher H;Newton ILG;Hardy RW
节肢动物内共生体Wolbachia pipientis是全球生物控制策略的一部分，以减少蚊子传播的RNA病毒如甲病毒的复制。我们以前证明了宿主胞嘧啶甲基转移酶，DNMT 2，在果蝇和病毒RNA作为病原体阻断过程中的细胞靶点的重要性。在这里，我们报告了DNMT 2在伊蚊物种中Wolbachia诱导的甲病毒抑制中的作用。病毒感染后，蚊子组织（包括唾液腺）中DNMT 2的表达升高。值得注意的是，这在沃尔巴克氏体定殖的动物中受到抑制，与病毒复制减少和子代病毒感染性降低一致。DNMT 2在培养的伊蚊细胞中的异位表达是前病毒，增加子代病毒的感染性，并且DNMT 2对病毒复制和感染性的这种作用依赖于其甲基转移酶活性。最后，通过LC-MS检查沃尔巴克氏体对病毒RNA修饰的影响，显示5-甲基胞嘧啶修饰量的减少与沃尔巴克氏体定殖的蚊子细胞和动物中DNMT 2的下调一致。总的来说，我们的研究结果支持这样的结论，即病毒RNA的5-甲基胞嘧啶修饰的破坏是病原体阻断的重要机制。这些数据还强调了表转录组修饰在调节基本甲病毒复制和传播过程中的重要作用。甲病毒是蚊媒病毒，是全球人类和牲畜发病率和死亡率的重要来源。减少这些病毒和相关病毒传播的干预措施包括将Wolbachia pipientis定殖蚊子引入病毒传播水平高的地理区域。沃尔巴克氏体是一种细胞内细菌，其在蚊子细胞中的存在导致抑制具有正义RNA基因组的病毒（例如甲病毒和黄病毒）的复制，该过程被称为“病原体阻断”。然而，在沃尔巴克氏体存在下抑制病毒复制的机制知之甚少。在这里，我们表明，沃尔巴克氏体导致宿主细胞胞嘧啶甲基转移酶的表达变化，导致病毒基因组RNA的甲基化的变化。次优甲基化降低了蚊子和哺乳动物宿主细胞中子代病毒的复制和感染性。沃尔巴克氏体的抑制作用可以通过恢复宿主甲基转移酶的表达来抑制，这表明它在沃尔巴克氏体介导的病原体阻断中起关键作用。此外，我们的研究结果表明，独立于沃尔巴克氏体的影响，病毒基因组的甲基化状态在病毒复制和传播的效率中起着重要的调节作用。
Arthropod endosymbiont Wolbachia pipientis is part of a global biocontrol strategy to reduce the replication of mosquito-borne RNA viruses such as alphaviruses. We previously demonstrated the importance of a host cytosine methyltransferase, DNMT2, in Drosophila and viral RNA as a cellular target during pathogen-blocking. Here we report a role for DNMT2 in Wolbachia-induced alphavirus inhibition in Aedes species. Expression of DNMT2 in mosquito tissues, including the salivary glands, is elevated upon virus infection. Notably, this is suppressed in Wolbachia-colonized animals, coincident with reduced virus replication and decreased infectivity of progeny virus. Ectopic expression of DNMT2 in cultured Aedes cells is proviral, increasing progeny virus infectivity, and this effect of DNMT2 on virus replication and infectivity is dependent on its methyltransferase activity. Finally, examining the effects of Wolbachia on modifications of viral RNA by LC-MS show a decrease in the amount of 5-methylcytosine modification consistent with the down-regulation of DNMT2 in Wolbachia colonized mosquito cells and animals. Collectively, our findings support the conclusion that disruption of 5-methylcytosine modification of viral RNA is a vital mechanism operative in pathogen blocking. These data also emphasize the essential role of epitranscriptomic modifications in regulating fundamental alphavirus replication and transmission processes. Alphaviruses are mosquito-transmitted viruses and a significant source of morbidity and mortality in humans and livestock worldwide. Interventions to reduce transmission of these and related viruses include the introduction of Wolbachia pipientis colonized mosquitoes into geographic regions with high levels of virus transmission. Wolbachia is an intracellular bacterium whose presence in mosquito cells leads to the inhibition of replication of viruses with positive-sense RNA genomes, such as alphaviruses and flaviviruses, a process referred to as “pathogen blocking”. However, the mechanisms by which virus replication is inhibited in the presence of Wolbachia are poorly understood. Here we show that Wolbachia leads to changes in the expression of a host cell cytosine methyltransferase resulting in changes in the methylation of the viral genomic RNA. Suboptimal methylation decreases replication and the infectivity of progeny viruses in mosquito and mammalian host cells. The inhibitory effects of Wolbachia can be suppressed by restoring expression of the host methyltransferase demonstrating that it plays a critical role in Wolbachia-mediated pathogen blocking. Further, our findings indicate that, independent of the effect of Wolbachia, the methylation state of the viral genome plays an important regulatory role in the efficiency of virus replication and transmission.
登录
查看更多内容
影响因子：
30.3
作者：
Kaur R;Shropshire JD;Cross KL;Leigh B;Mansueto AJ;Stewart V;Bordenstein SR;Bordenstein SR
通讯作者：
Bordenstein SR
影响因子：
6.4
作者：
Durbin AF;Wang C;Marcotrigiano J;Gehrke L
通讯作者：
Gehrke L
影响因子：
2.6
作者：
Diaz-Sanchez, S.;Hernandez-Jarguin, A.;de la Fuente, J.
通讯作者：
de la Fuente, J.
影响因子：
56.9
作者：
Goll, MG;Kirpekar, F;Bestor, TH
通讯作者：
Bestor, TH
影响因子：
5.3
作者：
Dunbar, Carmen A.;Rayaprolu, Vamseedhar;Jarrold, Martin F.
通讯作者：
Jarrold, Martin F.