Systematic and single cell analysis of Xenopus Piwi-interacting RNAs and Xiwi
Systematic and single cell analysis of Xenopus Piwi-interacting RNAs and Xiwi
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DOI：
10.1038/emboj.2009.237
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发表时间：
2009-10-07
期刊：
影响因子：
11.4
通讯作者：
Blower, Michael D.
中科院分区：
文献类型：
--
作者：
Lau, Nelson C.;Ohsumi, Toshiro;Blower, Michael D.
Piwi蛋白和Piwi相互作用RNA（piRNA）对于生殖细胞发育是必需的，但在大多数动物生殖细胞中，这些核糖核蛋白的分子机制的分析仍然具有挑战性。为了解决这一挑战，我们系统地表征了Xiwi（非洲爪蟾Piwi同源物）以及来自非洲爪蟾卵和卵母细胞的piRNA。我们使用大尺寸的非洲爪蟾卵在单细胞水平上分析小RNA，并在热带爪蟾基因组中发现了丰富的piRNA和大皮尔纳簇，其中一些类似于果蝇产生piRNA的弗拉门戈基因座。虽然大多数皮尔纳簇在卵中同时表达，但个体青蛙显示出不同的簇表达谱。Xiwi与微管和减数分裂纺锤体相关，并定位于生殖细胞形成的细胞质决定因素-生殖细胞。Xiwi以RNA依赖的方式与翻译调节因子结合，但爪蟾与Xiwi的相互作用不依赖于RNA。我们的研究通过显示个体动物可以具有不同的皮尔纳表达谱来增加对皮尔纳转录调控的了解。我们认为，除了调节转座因子，Xiwi可能在脊椎动物卵母细胞中指定RNA定位的功能。The EMBO Journal（2009）28，2945-2958. doi：10.1038/doj.2009.237; 2009年8月27日在线发布
Piwi proteins and Piwi-interacting RNAs (piRNAs) are essential for germ cell development, but analysis of the molecular mechanisms of these ribonucleoproteins remains challenging in most animal germ cells. To address this challenge, we systematically characterized Xiwi, a Xenopus Piwi homologue, and piRNAs from Xenopus eggs and oocytes. We used the large size of Xenopus eggs to analyze small RNAs at the single cell level, and find abundant piRNAs and large piRNA clusters in the Xenopus tropicalis genome, some of which resemble the Drosophila piRNA-generating flamenco locus. Although most piRNA clusters are expressed simultaneously in an egg, individual frogs show distinct profiles of cluster expression. Xiwi is associated with microtubules and the meiotic spindle, and is localized to the germ plasm-a cytoplasmic determinant of germ cell formation. Xiwi associates with translational regulators in an RNA-dependent manner, but Xenopus tudor interacts with Xiwi independently of RNA. Our study adds insight to piRNA transcription regulation by showing that individual animals can have differential piRNA expression profiles. We suggest that in addition to regulating transposable elements, Xiwi may function in specifying RNA localization in vertebrate oocytes. The EMBO Journal (2009) 28, 2945-2958. doi: 10.1038/emboj.2009.237; Published online 27 August 2009