Identification of eight proteins that cross-link to pre-mRNA in the yeast commitment complex.
Identification of eight proteins that cross-link to pre-mRNA in the yeast commitment complex.
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DOI：
10.1101/gad.13.5.581
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发表时间：
1999-03
影响因子：
10.5
通讯作者：
D. Zhang;M. Rosbash
中科院分区：
文献类型：
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作者：
D. Zhang;M. Rosbash
在酵母承诺复合体和哺乳动物E复合体中，U1 snRNA的5′端与pre-mRNA保守的5′剪接位点区域之间存在重要的碱基配对相互作用。但在早期剪接复合体的这一区域或附近，没有确定剪接蛋白和前mrna底物之间的蛋白质接触。为了解决这个问题，我们使用4-硫脲取代的含有5'剪切位点的rna作为底物，并鉴定了8种交联蛋白，所有这些蛋白之前都被鉴定为承诺复合物组分。这些蛋白定位于三个结构域：外显子、5' s区的六个核苷酸和下游的内含子。结果表明，承诺复合体的5'剪接位点区域和周围有密集的蛋白质接触，其中一些对U1 snRNP-pre-mRNA复合体的形成或稳定性有重要贡献。
In the yeast commitment complex and the mammalian E complex, there is an important base-pairing interaction between the 5' end of U1 snRNA and the conserved 5' splice site region of pre-mRNA. But no protein contacts between splicing proteins and the pre-mRNA substrate have been defined in or near this region of early splicing complexes. To address this issue, we used 4-thiouridine-substituted 5' splice site-containing RNAs as substrates and identified eight cross-linked proteins, all of which were identified previously as commitment complex components. The proteins were localized to three domains: the exon, the six nucleotides of the 5' ss region, and the downstream intron. The results indicate that the 5' splice site region and environs are dense with protein contacts in the commitment complex and suggest that some of them make important contributions to formation or stability of the U1 snRNP-pre-mRNA complex.