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首页> 外文期刊>Molecular and Cellular Biology >Recognition of mutant and cryptic 5' splice sites by the U1 small nuclear ribonucleoprotein in vitro.
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Recognition of mutant and cryptic 5' splice sites by the U1 small nuclear ribonucleoprotein in vitro.

机译：U1小核糖核糖核蛋白在体外识别突变和隐性5'剪接位点。

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摘要

We examined the ability of U1 small nuclear ribonucleoproteins (U1 snRNPs) to recognize mutant and cryptic 5' splice sites on beta-globin pre-mRNA substrates using an RNase T1 protection assay. When U1 snRNPs were prebound to anti-(U1)RNP antibodies, we detected binding to mutant but not to cryptic 5' splice sites on several substrates. By contrast, in a splicing extract at 0 degree C, neither the mutated nor cryptic 5' splice sites of a human beta-globin transcript were selected as protected fragments with the same antibodies. However, after incubation of the transcript in the extract to yield splicing intermediates, fragments that included a cryptic 5' splice site were detected. The results of our analyses suggest that U1 snRNPs are involved in recognizing cryptic 5' splice sites but that interactions with other splicing components are required to stabilize the association.
机译：我们检查了使用核糖核酸酶T1保护测定法的U1小核糖核蛋白（U1 snRNPs）识别β-珠蛋白pre-mRNA底物上的突变和隐性5'剪接位点的能力。当U1 snRNPs预结合抗（U1）RNP抗体时，我们检测到与突变体的结合，但未结合几种底物上的隐性5'剪接位点。相反，在0℃的剪接提取物中，人β-珠蛋白转录物的突变的或隐含的5'剪接位点均未被选择为具有相同抗体的受保护片段。然而，在提取物中温育转录物以产生剪接中间体之后，检测到包括隐蔽的5'剪接位点的片段。我们的分析结果表明，U1 snRNPs参与识别隐蔽的5'剪接位点，但需要与其他剪接组件进行相互作用以稳定该关联。

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