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首页> 外文期刊>Molecular and Cellular Biology >SR proteins promote the first specific recognition of Pre-mRNA and are present together with the U1 small nuclear ribonucleoprotein particle in a general splicing enhancer complex.
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SR proteins promote the first specific recognition of Pre-mRNA and are present together with the U1 small nuclear ribonucleoprotein particle in a general splicing enhancer complex.

机译：SR蛋白促进Pre-mRNA的第一个特异性识别，并与U1小核糖核蛋白颗粒一起存在于一般的剪接增强子复合物中。

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We show that addition of SR proteins to in vitro splicing extracts results in a significant increase in assembly of the earliest prespliceosomal complex E and a corresponding decrease in assembly of the heterogeneous nuclear ribonucleoprotein (hnRNP) complex H. In addition, SR proteins promote formation of the E5' and E3' complexes that assemble on RNAs containing only 5' and 3' splice sites, respectively. We conclude that SR proteins promote the earliest specific recognition of both the 5' and 3' splice sites and are limiting for this function in HeLa nuclear extracts. Using UV cross-linking, we demonstrate specific, splice site-dependent RNA-protein interactions of SR proteins in the E, E5', and E3' complexes. SR proteins do not UV cross-link in the H complex, and conversely, hnRNP cross-linking is largely excluded from the E-type complexes. We also show that a discrete complex resembling the E5' complex assembles on both purine-rich and non-purine-rich exonic splicing enhancers. This complex, which we have designated the Enhancer complex, contains U1 small nuclear RNP (snRNP) and is associated with different SR protein family members, depending on the sequence of the enhancer. We propose that both downstream 5' splice site enhancers and exonic enhancers function by establishing a network of pre-mRNA-protein and protein-protein interactions involving U1 snRNP, SR proteins, and U2AF that is similar to the interactions that bring the 5' and 3' splice sites together in the E complex.
机译：我们表明，在体外剪接提取物中加入SR蛋白会导致最早的剪枝前体复合体E的装配显着增加，而异质核糖核蛋白（hnRNP）复杂H的装配也将相应减少。 E5'和E3'复合物分别在仅包含5'和3'剪接位点的RNA上组装。我们得出的结论是，SR蛋白促进5'和3'剪接位点的最早的特异性识别，并且在HeLa核提取物中限制了此功能。使用UV交联，我们证明了E，E5'和E3'复合物中SR蛋白的特异性，剪接位点依赖性RNA-蛋白相互作用。 SR蛋白在H络合物中不发生UV交联，相反，hnRNP交联在E型络合物中被大大排除。我们还显示，类似于E5'复合物的离散复合物在富含嘌呤和非富含嘌呤的外显子剪接增强子上组装。我们将这种复合物称为增强子复合物，它包含U1小核RNP（snRNP），并与不同的SR蛋白家族成员相关，具​​体取决于增强子的序列。我们建议下游5'剪接位点增强子和外显子增强子通过建立涉及m1蛋白，蛋白和蛋白相互作用的网络（涉及U1 snRNP，SR蛋白和U2AF）来起作用，这类似于带来5'和3'剪接位点在E复合物中。

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