RNA结合蛋白HYL1和SE促进DCL1对初级微小RNA（pri-miRNA）进行精确的体外加工。
The RNA-binding proteins HYL1 and SE promote accurate in vitro processing of pri-miRNA by DCL1.
作者信息
Dong Zhicheng, Han Meng-Hsuan, Fedoroff Nina
机构信息
Biology Department and Huck Institutes of Life Sciences, Pennsylvania State University, University Park, PA 16802, USA.
出版信息
Proc Natl Acad Sci U S A. 2008 Jul 22;105(29):9970-5. doi: 10.1073/pnas.0803356105. Epub 2008 Jul 16.
Abstract
The results of genetic studies in Arabidopsis indicate that three proteins, the RNase III DICER-Like1 (DCL1), the dsRNA-binding protein HYPONASTIC LEAVES1 (HYL1), and the C2H2 Zn-finger protein SERRATE (SE), are required for the accurate processing of microRNA (miRNA) precursors in the plant cell nucleus. To elucidate the biochemical mechanism of miRNA processing, we developed an in vitro miRNA processing assay using purified recombinant proteins. We find that DCL1 alone releases 21-nt short RNAs from dsRNA as well as synthetic miR167b precursor RNAs. However, correctly processed miRNAs constitute a minority of the cleavage products. We show that recombinant HYL1 and SE proteins accelerate the rate of DCL1-mediated cleavage of pre- and pri-miR167b substrates and promote accurate processing.
摘要
拟南芥的遗传学研究结果表明，植物细胞核中微小RNA（miRNA）前体的精确加工需要三种蛋白质，即核糖核酸酶III类Dicer样蛋白1（DCL1）、双链RNA结合蛋白叶片下卷1（HYL1）和C2H2锌指蛋白锯齿状蛋白（SE）。为阐明miRNA加工的生化机制，我们利用纯化的重组蛋白开发了一种体外miRNA加工检测方法。我们发现，单独的DCL1能从双链RNA以及合成的miR167b前体RNA中释放出21个核苷酸的短RNA。然而，正确加工的miRNA仅占切割产物的少数。我们表明，重组HYL1和SE蛋白可加速DCL1介导的前体miR167b和初级miR167b底物的切割速率，并促进精确加工。
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