DYNAMIC ASSOCIATION OF PROTEINS WITH THE PRE-MESSENGER-RNA BRANCH REGION
DYNAMIC ASSOCIATION OF PROTEINS WITH THE PRE-MESSENGER-RNA BRANCH REGION
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DOI：
10.1101/gad.8.24.3008
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发表时间：
1994-12-15
影响因子：
10.5
通讯作者：
SHARP, PA
中科院分区：
文献类型：
--
作者：
MACMILLAN, AM;QUERY, CC;SHARP, PA
在前体mRNA剪接过程中，蛋白质与分支位点区域的关联使用一种新的方法来探测，该方法利用光试剂二苯甲酮对前体mRNA进行位点特异性修饰。三组蛋白质的区别与前mRNA的动力学，其协会与离散剪接复合物，并通过其不同的因素的要求。早期U1 snRNP依赖性的分支区域与p80物质的交联，随后是与U2 snRNP-前mRNA复合物相关的p14、p35和p150多肽的交联。伴随着剪接体的形成，分支区域的蛋白因子发生重排，其中p35和p150交联被p220和p70种类取代。这些结果表明，分支区域在剪接体组装过程中被多种不同的蛋白质以动态方式识别。
The association of proteins with the branch site region during pre-mRNA splicing was probed using a novel methodology to site-specifically modify the pre-mRNA with the photo-reagent benzophenone. Three sets of proteins were distinguished by the kinetics of their associations with pre-mRNAs, by their association with discrete splicing complexes, and by their differing factor requirements. An early U1 snRNP-dependent cross-link of the branch region to a p80 species was followed by cross-links to p14, p35, and p150 polypeptides associated with the U2 snRNP-pre-mRNA complex. Concomitant with formation of the spliceosome, a rearrangement of protein factors about the branch region occurred, in which the p35 and p150 cross-links were replaced by p220 and p70 species. These results establish that the branch region is recognized in a dynamic fashion by multiple distinct proteins during the course of spliceosomal assembly.