Division of Biological and Environmental Sciences and Engineering, King Abdullah University of Science and Technology, Thuwal 23955-6900, Kingdom of Saudi Arabia and Center for Desert Agriculture, King Abdullah University of Science and Technology, Thuwal 23955-6900, Kingdom of Saudi Arabia.
Nucleic Acids Res. 2013 Oct;41(19):9129-40. doi: 10.1093/nar/gkt667. Epub 2013 Aug 5.
Efficient and precise microRNA (miRNA) biogenesis in Arabidopsis is mediated by the RNaseIII-family enzyme DICER-LIKE 1 (DCL1), double-stranded RNA-binding protein HYPONASTIC LEAVES 1 and the zinc-finger (ZnF) domain-containing protein SERRATE (SE). In the present study, we examined primary miRNA precursor (pri-miRNA) processing by highly purified recombinant DCL1 and SE proteins and found that SE is integral to pri-miRNA processing by DCL1. SE stimulates DCL1 cleavage of the pri-miRNA in an ionic strength-dependent manner. SE uses its N-terminal domain to bind to RNA and requires both N-terminal and ZnF domains to bind to DCL1. However, when DCL1 is bound to RNA, the interaction with the ZnF domain of SE becomes indispensible and stimulates the activity of DCL1 without requiring SE binding to RNA. Our results suggest that the interactions among SE, DCL1 and RNA are a potential point for regulating pri-miRNA processing.
拟南芥中高效精确的 microRNA (miRNA) 生物发生是由 RNaseIII 家族酶 DICER-LIKE 1 (DCL1)、双链 RNA 结合蛋白 HYPONASTIC LEAVES 1 和含锌指 (ZnF) 结构域的蛋白 SERRATE (SE) 介导的。在本研究中，我们使用高度纯化的重组 DCL1 和 SE 蛋白研究了初级 miRNA 前体 (pri-miRNA) 的加工，发现 SE 是 DCL1 加工 pri-miRNA 的必需成分。SE 以离子强度依赖的方式刺激 DCL1 对 pri-miRNA 的切割。SE 使用其 N 端结构域结合 RNA，并需要 N 端和 ZnF 结构域结合 DCL1。然而，当 DCL1 与 RNA 结合时，与 SE 的 ZnF 结构域的相互作用变得不可或缺，并在不需要 SE 与 RNA 结合的情况下刺激 DCL1 的活性。我们的结果表明，SE、DCL1 和 RNA 之间的相互作用是调节 pri-miRNA 加工的一个潜在点。
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