Center for Plant Biology, School of Life Sciences, Tsinghua University, Beijing, China.
Tsinghua-Peking Center for Life Sciences, Beijing, China.
Nat Cell Biol. 2021 Jan;23(1):32-39. doi: 10.1038/s41556-020-00606-5. Epub 2020 Dec 7.
MicroRNA (miRNA) production entails the step-wise processing of primary miRNAs (pri-miRNAs) into precursor miRNAs (pre-miRNAs) and miRNA/* duplexes by Dicing complexes containing DCL1, HYL1 and SE, which are localized in nuclear dicing bodies (D-bodies). Here, we show that D-bodies are phase-separated condensates. SE forms droplets and drives DCL1, HYL1 and pri/pre-miRNAs into the droplets in vitro, and mutation of SE abrogates the formation of D-bodies in vivo, which indicates that D-bodies arise through SE-mediated phase separation. Disruption of SE phase separation greatly reduces its activity in promoting miRNA processing both in vitro and in vivo. We further show that pre-miRNAs are processed into miRNA/* duplexes in the droplets and, after processing, miRNA/* duplexes are bound by HYL1 and released from the droplets. Our findings provide evidence that efficient miRNA processing depends on the SE-phase-separation-mediated formation of D-bodies and suggest a paradigm that the products made in phase-separated condensates can be shipped out for subsequent processes.
MicroRNA (miRNA) 的生成需要 Dicer 复合物逐步加工初级 miRNA (pri-miRNA) 成为前体 miRNA (pre-miRNA) 和 miRNA/*双链,该复合物包含 DCL1、HYL1 和 SE,定位于核 Dicer 体 (D-body) 中。在这里,我们表明 D-body 是相分离的凝聚物。SE 形成液滴,并在体外将 DCL1、HYL1 和 pri/pre-miRNA 驱动到液滴中,而 SE 的突变会在体内破坏 D-body 的形成,这表明 D-body 是通过 SE 介导的相分离形成的。SE 相分离的破坏极大地降低了其在体外和体内促进 miRNA 加工的活性。我们进一步表明,pre-miRNA 在液滴中被加工成 miRNA/*双链,并且在加工后,miRNA/*双链被 HYL1 结合并从液滴中释放出来。我们的发现提供了证据表明有效的 miRNA 加工取决于 SE 相分离介导的 D-body 的形成,并提出了一个范例,即在相分离凝聚物中产生的产物可以被运出以供后续处理。