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Structure and function of Zucchini endoribonuclease in piRNA biogenesis

机译：西葫芦内切核糖核酸酶在piRNA生物发生中的结构和功能

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摘要

PlWI-interacting RNAs (piRNAs) silence transposons to maintain genome integrity in animal germ lines. piRNAs are classified as primary and secondary piRNAs, depending on their biogenesis machinery. Primary piRNAs are processed from long non-coding RNA precursors transcribed from piRNA clusters in the genome through the primary processing pathway. Although the existence of a ribonuclease participating in this pathway has been predicted, its molecular identity remained unknown. Here we show that Zucchini (Zuc), a mitochondrial phospholipase D (PLD) superfamily member is an endoribonuclease essential for primary piRNA biogenesis. We solved the crystal structure of Drosophila melanogaster Zuc (DmZuc) at 1.75 A resolution. The structure revealed that DmZuc has a positively charged, narrow catalytic groove at the dimer interface, which could accommodate a single-stranded, but not a double-stranded, RNA. DmZuc and the mouse homologue MmZuc (also known as Pld6 and MitoPLD)12"14 showed endoribonuclease activity for single-stranded RNAs in vitro. The RNA cleavage products bear a 5'-monophosphate group, a hallmark of mature piRNAs. Mutational analyses revealed that the conserved active-site residues of DmZuc are critical for the ribonuclease activity in vitro, and for piRNA maturation and transposon silencing in vivo. We propose a model for piRNA biogenesis in animal germ lines, in which the Zuc endoribonuclease has a key role in primary piRNA maturation.
机译：与PlWI相互作用的RNA（piRNA）使转座子沉默，以维持动物种系中的基因组完整性。根据其生物发生机制，将piRNA分为主要和次要piRNA。初级piRNA是通过初级加工途径从基因组中piRNA簇转录而来的长非编码RNA前体加工而成的。尽管已经预测了参与该途径的核糖核酸酶的存在，但其分子身份仍然未知。在这里，我们显示线粒体磷脂酶D（PLD）超家族成员西葫芦（Zuc）是主要piRNA生物发生所必需的内切核糖核酸酶。我们以1.75 A的分辨率解决了果蝇Zuc（DmZuc）的晶体结构。该结构表明，DmZuc在二聚体界面处具有带正电的狭窄催化槽，可以容纳单链而不是双链RNA。 DmZuc和小鼠同源物MmZuc（也称为Pld6和MitoPLD）12“ 14在体外显示出对单链RNA的核糖核酸内切酶活性。RNA裂解产物带有5'-单磷酸基团，这是成熟piRNA的标志。突变分析表明DmZuc保守的活性位点残基对于体外核糖核酸酶活性，体内piRNA成熟和转座子沉默至关重要，我们提出了动物胚系中piRNA生物发生的模型，其中Zuc内切核糖核酸酶在原核生物中起关键作用piRNA成熟。

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  • 来源
    《Nature》 |2012年第7423期|p.284-287|共4页
  • 作者

    Hiroshi Nishimasu; Hirotsugu Ishizu; Kuniaki Saito; Satoshi Fukuhara; Miharu K. Kamatani; Luc Bonnefond; Naoki Matsumoto; Tomohiro Nishizawa; Keita Nakanaga; Junken Aoki; Ryuichiro Ishitani; Haruhiko Siomi; Mikiko C. Siomi; Osamu Nureki;

  • 作者单位

    Department of Biophysics and Biochemistry, Graduate School of Science, The University of Tokyo, Tokyo 113-0032, Japan;

    Department of Biophysics and Biochemistry, Graduate School of Science, The University of Tokyo, Tokyo 113-0032, Japan,Department of Molecular Biology, Keio University School of Medicine, Tokyo 160-8582, Japan;

    Department of Molecular Biology, Keio University School of Medicine, Tokyo 160-8582, Japan;

    Department of Biophysics and Biochemistry, Graduate School of Science, The University of Tokyo, Tokyo 113-0032, Japan;

    Department of Molecular Biology, Keio University School of Medicine, Tokyo 160-8582, Japan;

    Department of Biophysics and Biochemistry, Graduate School of Science, The University of Tokyo, Tokyo 113-0032, Japan,Institut de Genetique et de Biologie Moleculaire et Cellulaire, UMR7104 CNRS-UDS, INSERM U964,1 rue Laurent Fries, BP 10142,67404 Illkirch Cedex, France;

    Department of Biophysics and Biochemistry, Graduate School of Science, The University of Tokyo, Tokyo 113-0032, Japan;

    Department of Biophysics and Biochemistry, Graduate School of Science, The University of Tokyo, Tokyo 113-0032, Japan;

    Graduate School of Pharmaceutical Sciences, Tohoku University, Miyagi 980-8578, Japan;

    Graduate School of Pharmaceutical Sciences, Tohoku University, Miyagi 980-8578, Japan;

    Department of Biophysics and Biochemistry, Graduate School of Science, The University of Tokyo, Tokyo 113-0032, Japan;

    Department of Molecular Biology, Keio University School of Medicine, Tokyo 160-8582, Japan;

    Department of Biophysics and Biochemistry, Graduate School of Science, The University of Tokyo, Tokyo 113-0032, Japan,Department of Molecular Biology, Keio University School of Medicine, Tokyo 160-8582, Japan,Japan Science and Technology Agency (JST), Core Research for Evolutional Science and Technology (CREST), Saitama 332-0012, Japan;

    Department of Biophysics and Biochemistry, Graduate School of Science, The University of Tokyo, Tokyo 113-0032, Japan,Japan Science and Technology Agency (JST), Core Research for Evolutional Science and Technology (CREST), Saitama 332-0012, 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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