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Hierarchical roles of mitochondrial Papi and Zucchini in Bombyx germline piRNA biogenesis

机译：线粒体Papi和西葫芦在Bombyx种系piRNA生物发生中的分层作用

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摘要

PIWI-interacting RNAs (piRNAs) are small regulatory RNAs that bind to PIWI proteins to control transposons and maintain genome integrity in animal germ lines1-3. piRNA 3' end formation in the silkworm Bombyx mori has been shown to be mediated by the 3'-to-5' exonuclease Trimmer (Trim; known as PNLDC1 in mammals) 4, and piRNA intermediates are bound with PIWI anchored onto mitochondrial Tudor domain protein Papi(5). However, it remains unclear whether the Zucchini (Zuc) endonuclease and Nibbler (Nbr) 3'-to-5' exonuclease, both of which have pivotal roles in piRNA biogenesis in Drosophila(6-8), are required for piRNA processing in other species. Here we show that the loss of Zuc in Bombyx had no effect on the levels of Trim and Nbr, but resulted in the aberrant accumulation of piRNA intermediates within the Papi complex, and that these were processed to form mature piRNAs by recombinant Zuc. Papi exerted its RNA-binding activity only when bound with PIWI and phosphorylated, suggesting that complex assembly involves a hierarchical process. Both the 5' and 3' ends of piRNA intermediates within the Papi complex showed hallmarks of PIWI 'slicer' activity, yet no phasing pattern was observed in mature piRNAs. The loss of Zuc did not affect the 5'-and3'-end formation of the intermediates, strongly supporting the idea that the 5' end of Bombyx piRNA is formed by PIWI slicer activity, but independently of Zuc, whereas the 3' end is formed by the Zuc endonuclease. The Bombyx piRNA biogenesis machinery is simpler than that of Drosophila, because Bombyx has no transcriptional silencing machinery that relies on phased piRNAs.
机译：PIWI相互作用RNA（piRNA）是小的调节性RNA，与PIWI蛋白结合以控制转座子并维持动物种系1-3中的基因组完整性。已显示家蚕中piRNA 3'末端的形成是由3'至5'核酸外切酶Trimmer（Trim；在哺乳动物中称为PNLDC1）4介导的，并且piRNA中间体与锚定在线粒体Tudor域上的PIWI结合。蛋白Papi（5）。然而，尚不清楚西葫芦（Zuc）核酸内切酶和Nibbler（Nbr）3'至5'核酸外切酶，两者在果蝇（6-8）的piRNA生物发生中都起着关键作用，是否需要其他方法来进行piRNA加工。种类。在这里，我们显示了Bombyx中Zuc的丢失对Trim和Nbr的水平没有影响，但是导致piRNA中间产物在Papi复合物中异常积累，并且这些被重组Zuc加工成成熟的piRNA。 Papi仅在与PIWI结合并被磷酸化时才发挥其RNA结合活性，这表明复杂的组装过程涉及一个分层过程。 Papi复合物中的piRNA中间体的5'和3'末端均显示PIWI“切片器”活性的标志，但在成熟piRNA中未观察到定相模式。 Zuc的丢失并不影响中间体的5'和3'端形成，这强烈支持了Bombyx piRNA的5'端由PIWI切片机活性形成，但独立于Zuc的观点，而3'端则是由Zuc核酸内切酶形成。 Bombyx piRNA生物发生机制比果蝇更简单，因为Bombyx没有依赖相控piRNA的转录沉默机制。

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  • 来源
    《Nature》 |2018年第7695期|260-264|共5页
  • 作者

    Nishida Kazumichi M.; Sakakibara Kazuhiro; Iwasaki Yuka W.; Yamada Hiromi; Murakami Ryo; Murota Yukiko; Kawamura Takeshi; Kodama Tatsuhiko; Siomi Haruhiko; Siomi Mikiko C.;

  • 作者单位

    Univ Tokyo, Grad Sch Sci, Dept Biol Sci, Tokyo 1130032, Japan;

    Univ Tokyo, Grad Sch Sci, Dept Biol Sci, Tokyo 1130032, Japan;

    Keio Univ, Dept Mol Biol, Sch Med, Tokyo 1628582, Japan;

    Univ Tokyo, Grad Sch Sci, Dept Biol Sci, Tokyo 1130032, Japan;

    Univ Tokyo, Grad Sch Sci, Dept Biol Sci, Tokyo 1130032, Japan;

    Univ Tokyo, Grad Sch Sci, Dept Biol Sci, Tokyo 1130032, Japan;

    Univ Tokyo, Isotope Sci Ctr, Prote Lab, Tokyo 1130032, Japan;

    Univ Tokyo, Res Ctr Adv Sci & Technol, Lab Syst Biol & Med, Tokyo 1538904, Japan;

    Keio Univ, Dept Mol Biol, Sch Med, Tokyo 1628582, Japan;

    Univ Tokyo, Grad Sch Sci, Dept Biol Sci, Tokyo 1130032, Japan;

  • 收录信息 美国《科学引文索引》(SCI);美国《工程索引》(EI);美国《生物学医学文摘》(MEDLINE);美国《化学文摘》(CA);
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