The interaction between DCL1 and HYL1 is important for efficient and precise processing of pri-miRNA in plant microRNA biogenesis
The interaction between DCL1 and HYL1 is important for efficient and precise processing of pri-miRNA in plant microRNA biogenesis
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DOI：
10.1261/rna.2146906
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发表时间：
2006-02-01
期刊：
影响因子：
4.5
通讯作者：
Watanabe, Y
中科院分区：
文献类型：
--
作者：
Kurihara, Y;Takashi, Y;Watanabe, Y
据报道，一些双链RNA(DsRNA)结合蛋白与小RNA生物发生相关的RNaseIII酶相互作用。然而，人们对它们的生物学意义知之甚少。在这里，我们研究了拟南芥microRNA-(MiRNA)产生酶DCL1和dsRNA结合蛋白HYL1之间的关系。在hyi1-2突变体中，miR163的生物发生加工步骤部分受损，pri-miR163的积累增加，短的Pre-miR163和成熟的miR163的积累减少，以及Pri-miR163茎结构中错位的裂解。这些错位的切割类似于之前在dcl1-9突变体中观察到的，其中蛋白质的第二个双链RNA结合域被破坏。利用农杆菌介导法在烟草中瞬时表达的免疫沉淀实验表明，HYL1能够与野生型DCL1蛋白形成复合体，但不能与突变型DCL1-9蛋白形成复合体。我们还研究了hyl1-2和dcl1-9中的miR164b和miR166a的生物发生。Pri-miRNAs的积累增加，前miRNAs和成熟miRNAs的积累减少。仅在dcl1-9中观察到pri-miR164b上的错位切割，而在hyl1-2中未观察到错位切割，而在两个突变体中均未观察到pri-miR166a上的错位切割。这些结果表明，HYL1通过与DCL1的相互作用，帮助Pri-miRNA高效而精确地切割。
It has been reported that some double-stranded RNA (dsRNA) binding proteins interact with small RNA biogenesis-related RNase III enzymes. However, their biological significance is poorly understood. Here we examine the relationship between the Arabidopsis microRNA- (miRNA) producing enzyme DCL1 and the dsRNA binding protein HYL1. In the hyi1-2 mutant, the processing steps of miR163 biogenesis were partially impaired; increased accumulation of pri-miR163 and reduced accumulation of short pre-miR163 and mature miR163 as well as misplaced cleavages in the stem structure of pri-miR163 were detected. These misplaced cleavages were similar to those previously observed in the dcl1-9 mutant, in which the second double-stranded RNA binding domain of the protein was disrupted. An immunoprecipitation assay using Agrobacterium-mediated transient expression in Nicotiana benthamiana showed that HYL1 was able to form a complex with wild-type DCL1 protein, but not with the dcl1-9 mutant protein. We also examined miR164b and miR166a biogenesis in hyl1-2 and dcl1-9. increased accumulation of pri-miRNAs and reduced accumulation of pre-miRNAs and mature miRNAs were detected. Misplaced cleavage on pri-miR164b was observed only in dcl1-9 but not in hyl1-2, whereas not on pri-miR166a in either mutant. These results indicate that HYL1 has a function in assisting efficient and precise cleavage of pri-miRNA through interaction with DCL1.