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首页> 外文期刊>Molecular and Cellular Biology >Short artificial hairpins sequester splicing signals and inhibit yeast pre-mRNA splicing.
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Short artificial hairpins sequester splicing signals and inhibit yeast pre-mRNA splicing.

机译：短的人造发夹螯合剪接信号并抑制酵母前mRNA剪接。

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To examine the stability of yeast (Saccharomyces cerevisiae) pre-mRNA structures, we inserted a series of small sequence elements that generated potential RNA hairpins at the 5' splice site and branch point regions. We analyzed spliceosome assembly and splicing in vitro as well as splicing and nuclear pre-mRNA retention in vivo. Surprisingly, the inhibition of in vivo splicing approximately paralleled that of in vitro splicing. Even a 6-nucleotide hairpin could be shown to inhibit splicing, and a 15-nucleotide hairpin gave rise to almost complete inhibition. The in vitro results indicate that hairpins that sequester the 5' splice site have a major effect on the early steps of spliceosome assembly, including U1 small nuclear ribonucleoprotein binding. The in vivo experiments lead to comparable conclusions as the sequestering hairpins apparently result in the transport of pre-mRNA to the cytoplasm. The observations are compared with previous data from both yeast and mammalian systems and suggest an important effect of pre-mRNA structure on in vivo splicing.
机译：为了检查酵母（Saccharomyces cerevisiae）pre-mRNA结构的稳定性，我们插入了一系列小序列元件，这些元件在5'剪接位点和分支点区域产生了潜在的RNA发夹。我们分析了剪接体组装和体外剪接以及体内剪接和核前mRNA保留。令人惊讶的是，体内剪​​接的抑制与体外剪接的抑制近似平行。甚至可以显示6个核苷酸的发夹会抑制剪接，而15个核苷酸的发夹会产生几乎完全的抑制作用。体外结果表明，隔离5'剪接位点的发夹对剪接体组装的早期步骤（包括U1小核糖核糖核蛋白结合）具有重大影响。体内实验得出了可比的结论，因为螯合的发夹显然导致前mRNA转运到细胞质。将这些观察结果与来自酵母和哺乳动物系统的先前数据进行了比较，并表明前mRNA结构对体内剪接具有重要作用。

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