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Endogenous piRNA-guided slicing triggers responder and trailer piRNA production from viral RNA in Aedes aegypti mosquitoes
Nucleic Acids Research ( IF 16.6 ) Pub Date : 2021-07-20 , DOI: 10.1093/nar/gkab640
Joep Joosten 1 , Gijs J Overheul 1 , Ronald P Van Rij 1 , Pascal Miesen 1
Affiliation  

In the germline of animals, PIWI interacting (pi)RNAs protect the genome against the detrimental effects of transposon mobilization. In Drosophila, piRNA-mediated cleavage of transposon RNA triggers the production of responder piRNAs via ping-pong amplification. Responder piRNA 3′ end formation by the nuclease Zucchini is coupled to the production of downstream trailer piRNAs, expanding the repertoire of transposon piRNA sequences. In Aedes aegypti mosquitoes, piRNAs are generated from viral RNA, yet, it is unknown how viral piRNA 3′ ends are formed and whether viral RNA cleavage gives rise to trailer piRNA production. Here we report that in Ae. aegypti, virus- and transposon-derived piRNAs have sharp 3′ ends, and are biased for downstream uridine residues, features reminiscent of Zucchini cleavage of precursor piRNAs in Drosophila. We designed a reporter system to study viral piRNA 3′ end formation and found that targeting viral RNA by abundant endogenous piRNAs triggers the production of responder and trailer piRNAs. Using this reporter, we identified the Ae. aegypti orthologs of Zucchini and Nibbler, two nucleases involved in piRNA 3′ end formation. Our results furthermore suggest that autonomous piRNA production from viral RNA can be triggered and expanded by an initial cleavage event guided by genome-encoded piRNAs.

中文翻译:

内源性 piRNA 引导的切片触发埃及伊蚊中病毒 RNA 产生响应者和拖车 piRNA

在动物的生殖系中,PIWI 相互作用 (pi) RNA 保护基因组免受转座子动员的有害影响。在果蝇中,piRNA 介导的转座子 RNA 切割通过乒乓放大触发反应 piRNA 的产生。核酸酶 Zucchini 形成的反应器 piRNA 3' 末端与下游拖车 piRNA 的产生相结合,从而扩展了转座子 piRNA 序列的库。在埃及伊蚊中,piRNAs 是由病毒 RNA 产生的,但是,尚不清楚病毒 piRNA 3' 端是如何形成的,以及病毒 RNA 切割是否会导致尾 piRNA 产生。在这里,我们在 Ae 中报告了这一点。埃及伊蚊、病毒和转座子衍生的 piRNA 具有尖锐的 3' 末端,并且偏向于下游尿苷残基,其特征让人联想到西葫芦对果蝇中前体 piRNA 的切割。我们设计了一个报告系统来研究病毒 piRNA 3' 末端的形成,并发现通过丰富的内源性 piRNA 靶向病毒 RNA 会触发响应者和尾随 piRNA 的产生。使用这个记者,我们确定了 Ae。Zucchini 和 Nibbler 的 aegypti 直系同源物,这两种核酸酶参与 piRNA 3' 末端的形成。我们的研究结果进一步表明,病毒 RNA 的自主 piRNA 生产可以通过基因组编码的 piRNA 引导的初始切割事件来触发和扩展。
更新日期:2021-07-20
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