Kalmykova Alla I, Klenov Mikhail S, Gvozdev Vladimir A
Institute of Molecular Genetics RAS Kurchatov square 2, 123 182 Moscow, Russia.
Nucleic Acids Res. 2005 Apr 7;33(6):2052-9. doi: 10.1093/nar/gki323. Print 2005.
Proteins of the Argonaute family have been identified as key components of RNA interference (RNAi) pathway. RNAi-related mechanisms are implicated in the regulation of gene expression and repression of transposable elements in eukaryotes. The piwi gene encoding protein of the Drosophila Argonaute family was shown to be required for the germ stem cells maintenance. Here, we show that piwi is involved in silencing of LTR retrotransposons in testes. piwi mutations led to derepression of endogenous retrotransposon copia as well as to upregulation of the reporter gene driven by copia LTR. piwi mutation causes accumulation of retrotransposon mdg1 transcripts at the apical tip of testes, including germinal proliferative center where PIWI protein was shown to be expressed. We applied inverse PCR approach to detect the newly arisen insertions of the mdg1 retrotransposon in the progeny of individual piwi mutant males. Owing to piwi mutation a high rate of mdg1 transpositions was revealed. Thus, piwi is involved in the silencing of retrotransposons in the precursors of male gametes. Our results provide the first evidence that protein of the Argonaute family prevents retrotranspositions. It is supposed that the disturbance of RNA silencing system in germinal cells might cause transposition burst.
AGO蛋白家族的蛋白质已被鉴定为RNA干扰(RNAi)途径的关键组成部分。RNAi相关机制参与真核生物中基因表达的调控和转座元件的抑制。编码果蝇AGO蛋白家族蛋白质的piwi基因被证明是生殖干细胞维持所必需的。在此,我们表明piwi参与睾丸中LTR逆转座子的沉默。piwi突变导致内源性逆转座子copia的去抑制以及由copia LTR驱动的报告基因的上调。piwi突变导致逆转座子mdg1转录本在睾丸顶端积累,包括显示PIWI蛋白表达的生发增殖中心。我们应用反向PCR方法检测单个piwi突变雄性后代中mdg1逆转座子的新插入。由于piwi突变,揭示了mdg1转座的高发生率。因此,piwi参与雄配子前体中逆转座子的沉默。我们的结果提供了第一个证据,表明AGO蛋白家族的蛋白质可防止逆转座。据推测,生殖细胞中RNA沉默系统的紊乱可能导致转座爆发。