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首页> 外文期刊>Molecular cell >HnRNP A1 Proofreads 3' Splice Site Recognition by U2AF
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HnRNP A1 Proofreads 3' Splice Site Recognition by U2AF

机译：HnRNP A1通过U2AF校对3'剪接位点识别

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摘要

One of the earliest steps in metazoan pre-mRNA splicing involves binding of U2 snRNP auxiliary factor (U2AF) 65 KDa subunit to the polypyrimidine (Py) tract and of the 35 KDa subunit to the invariant AG dinucleotide at the intron 3' end. Here we use invitro and invivo depletion, as well as reconstitution assays using purified components, to identify hnRNP A1 as an RNA binding protein that allows U2AF todiscriminate between pyrimidine-rich RNA sequences followed or not by a 3' splice site AG. Biochemical and NMR data indicate that hnRNP A1 forms a ternary complex with the U2AF heterodimer on AG-containing/uridine-rich RNAs, while it displaces U2AF from non-AG-containing/uridine-rich RNAs, an activity that requires the glycine-rich domain of hnRNP A1. Consistent with the functional relevance of this activity for splicing, proofreading assays reveal a role for hnRNP A1 in U2AF-mediated recruitment of U2 snRNP to the pre-mRNA.
机译：后生动物mRNA剪接的最早步骤之一涉及将U2 snRNP辅助因子（U2AF）65 KDa亚基与聚嘧啶（Py）束结合，并将35 KDa亚基与内含子3'端的不变AG二核苷酸结合。在这里，我们使用体内和体内耗竭，以及使用纯化成分的重组测定法，将hnRNP A1鉴定为RNA结合蛋白，可让U2AF区分富含嘧啶的RNA序列，然后区分3'剪接位点AG。生化和NMR数据表明，hnRNP A1与含AG /富尿苷的RNA上的U2AF异二聚体形成三元复合物，同时将U2AF从不含AG的富尿苷的RNA上置换出来，这一活动需要富含甘氨酸的甘氨酸hnRNP A1的结构域。与该活性与剪接的功能相关性一致，校对分析揭示了hnRNP A1在U2AF介导的U2 snRNP募集至前mRNA中的作用。

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