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首页> 外文期刊>Molecular and Cellular Biology >U1 small nuclear ribonucleoproteins are required early during spliceosome assembly.
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U1 small nuclear ribonucleoproteins are required early during spliceosome assembly.

机译：在剪接体组装的早期需要U1小核糖核蛋白。

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U1 small nuclear ribonucleoproteins (snRNPs) are required for in vitro splicing of pre-mRNA. Sequences within U1 RNA hybridize to, and thus recognize, 5' splice junctions. We have investigated the mechanism of association of U1 snRNPs with the spliceosome. U1-specific antibodies detected U1 association with precursor RNA early during assembly. Removal of the 5' terminal sequences of U1 RNA by oligo-directed cleavage or removal of U1 snRNPs by immunoprecipitation prior to the addition of precursor RNA depressed the association of all snRNPs with precursor RNA as detected by immunoprecipitation of splicing complexes by either Sm or U1-specific antibodies. Assembly of the spliceosome as monitored by gel electrophoresis was also depressed after cleavage of U1 RNA. The dependency of Sm precipitability of precursor RNA upon the presence of U1 snRNPs suggests that U1 snRNPs participate in the early recognition of substrate RNAs by U2 to U6 snRNPs. Although removal of the 5'-terminal sequences of U1 depressed U1 snRNP association with precursor RNA, it did not eliminate it, suggesting semistable association of U1 snRNPs with the assembling spliceosome in the absence of U1 RNA hybridization. This association was not dependent upon 5' splice junction sequences but was dependent upon 3' intronic sequences, indicating that U1 snRNPs interact with factors recognizing 3' intronic sequences. Mutual dependence of 5' and 3' recognition factors suggests significant snRNP-snRNP communication during early assembly.
机译：U1小核糖核糖蛋白（snRNPs）是pre-mRNA的体外剪接所必需的。 U1 RNA中的序列与5'剪接点杂交并因此识别。我们研究了U1 snRNPs与剪接体的关联机制。 U1特异性抗体可在组装早期检测到U1与前体RNA的缔合。在添加前体RNA之前通过寡核苷酸定向切割或通过免疫沉淀去除U1 snRNP来去除U1 RNA的5'末端序列，从而抑制了所有snRNP与前体RNA的缔合，如Sm或U1对拼接复合体的免疫沉淀所检测到的。特异性抗体。在切割U1 RNA后，还抑制了通过凝胶电泳监测的剪接体的组装。前体RNA的Sm沉淀性对U1 snRNPs的依赖性表明，U1 snRNPs参与了U2到U6 snRNPs对底物RNA的早期识别。尽管去除U1的5'-末端序列抑制了U1 snRNP与前体RNA的结合，但并没有消除它，这表明在没有U1 RNA杂交的情况下，U1 snRNPs与组装的剪接体是半稳定的结合。该关联不依赖于5'剪接连接序列，而是依赖于3'内含子序列，表明U1 snRNP与识别3'内含子序列的因子相互作用。 5'和3'识别因子的相互依赖表明在早期组装过程中重要的snRNP-snRNP通信。

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