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首页> 外文期刊>Nucleic Acids Research >Regulation of pri-miRNA processing by the hnRNP-like protein AtGRP7 in Arabidopsis
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Regulation of pri-miRNA processing by the hnRNP-like protein AtGRP7 in Arabidopsis

机译：拟南芥中hnRNP样蛋白AtGRP7对pri-miRNA加工的调控。

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摘要

The hnRNP-like glycine-rich RNA-binding protein AtGRP7 regulates pre-mRNA splicing in Arabidopsis. Here we used small RNA-seq to show that AtGRP7 also affects the miRNA inventory. AtGRP7 overexpression caused a significant reduction in the level of 30 miRNAs and an increase for 14miRNAs with a minimum log(2) fold change of +/- 0.5. Overaccumulation of several pri-miRNAs including pri-miR398b, pri-miR398c, pri-miR172b, pri-miR159a and pri-miR390 at the expense of the mature miRNAs suggested that AtGRP7 affects pri-miRNA processing. Indeed, RNA immunoprecipitation revealed that AtGRP7 interacts with these pri-miRNAs in vivo. Mutation of an arginine in the RNA recognition motif abrogated in vivo binding and the effect on miRNA and pri-miRNA levels, indicating that AtGRP7 inhibits processing of these pri-miRNAs by direct binding. In contrast, pri-miRNAs of selected miRNAs that were elevated or not changed in response to high AtGRP7 levels were not bound in vivo. Reduced accumulation of miR390, an initiator of trans-acting small interfering RNA (ta-siRNA) formation, also led to lower TAS3 ta-siRNA levels and increased mRNA expression of the target AUXIN RESPONSE FACTOR4. Furthermore, AtGRP7 affected splicing of pri-miR172b and pri-miR162a. Thus, AtGRP7 is an hnRNP-like protein with a role in processing of pri-miRNAs in addition to its role in pre-mRNA splicing.
机译：hnRNP样富含甘氨酸的RNA结合蛋白AtGRP7调节拟南芥中的pre-mRNA剪接。在这里，我们使用小的RNA-seq来表明AtGRP7也影响miRNA的库存。 AtGRP7的过表达导致30 miRNA的水平显着降低，而14 miRNA的升高则导致log（2）最小变化为+/- 0.5。包括pri-miR398b，pri-miR398c，pri-miR172b，pri-miR159a和pri-miR390在内的几种pri-miRNA的过度积累表明，AtGRP7影响pri-miRNA的加工。实际上，RNA免疫沉淀显示AtGRP7在体内与这些pri-miRNA相互作用。 RNA识别基序中精氨酸的突变消除了体内结合以及对miRNA和pri-miRNA水平的影响，表明AtGRP7通过直接结合抑制了这些pri-miRNA的加工。相反，响应于高AtGRP7水平而升高或未改变的所选miRNA的pri-miRNA在体内没有结合。反式小分子干扰RNA（ta-siRNA）形成引发剂miR390的积累减少，也导致TAS3 ta-siRNA水平降低，靶标AUXIN反应因子4的mRNA表达增加。此外，AtGRP7影响pri-miR172b和pri-miR162a的剪接。因此，AtGRP7是一种hnRNP样蛋白，除了在pre-mRNA剪接中还起着加工pri-miRNA的作用。

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