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Identification of piRNA Binding Sites Reveals the Argonaute Regulatory Landscape of the C. elegans Germline.
Cell ( IF 45.5 ) Pub Date : 2018-Feb-22 , DOI: 10.1016/j.cell.2018.02.002
En-Zhi Shen 1 , Hao Chen 2 , Ahmet R Ozturk 1 , Shikui Tu 3 , Masaki Shirayama 4 , Wen Tang 1 , Yue-He Ding 1 , Si-Yuan Dai 1 , Zhiping Weng 5 , Craig C Mello 4
Affiliation  

piRNAs (Piwi-interacting small RNAs) engage Piwi Argonautes to silence transposons and promote fertility in animal germlines. Genetic and computational studies have suggested that C. elegans piRNAs tolerate mismatched pairing and in principle could target every transcript. Here we employ in vivo cross-linking to identify transcriptome-wide interactions between piRNAs and target RNAs. We show that piRNAs engage all germline mRNAs and that piRNA binding follows microRNA-like pairing rules. Targeting correlates better with binding energy than with piRNA abundance, suggesting that piRNA concentration does not limit targeting. In mRNAs silenced by piRNAs, secondary small RNAs accumulate at the center and ends of piRNA binding sites. In germline-expressed mRNAs, however, targeting by the CSR-1 Argonaute correlates with reduced piRNA binding density and suppression of piRNA-associated secondary small RNAs. Our findings reveal physiologically important and nuanced regulation of individual piRNA targets and provide evidence for a comprehensive post-transcriptional regulatory step in germline gene expression.

中文翻译:

piRNA 结合位点的鉴定揭示了秀丽隐杆线虫种系的 Argonaute 监管格局。

piRNA(与 Piwi 相互作用的小 RNA)与 Piwi Argonautes 合作以沉默转座子并促进动物生殖系的生育能力。遗传和计算研究表明,秀丽隐杆线虫 piRNA 可以容忍错配配对,并且原则上可以针对每个转录本。在这里,我们采用体内交联来识别 piRNA 和靶 RNA 之间的转录组范围的相互作用。我们表明 piRNA 与所有种系 mRNA 结合,并且 piRNA 结合遵循类似 microRNA 的配对规则。与 piRNA 丰度相比,靶向与结合能的相关性更好,这表明 piRNA 浓度不会限制靶向。在被 piRNA 沉默的 mRNA 中,次级小 RNA 在 piRNA 结合位点的中心和末端积累。然而,在种系表达的 mRNA 中,CSR-1 Argonaute 的靶向作用与降低 piRNA 结合密度和抑制 piRNA 相关的次级小 RNA 相关。我们的研究结果揭示了个体 piRNA 靶标在生理上重要且细致入微的调控,并为生殖系基因表达中的全面转录后调控步骤提供了证据。
更新日期:2018-02-15
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