The human splicing factor ASF/SF2 can specifically recognize pre-mRNA 5' splice sites.
The human splicing factor ASF/SF2 can specifically recognize pre-mRNA 5' splice sites.
复制标题
人剪接因子 ASF/SF2 可以特异性识别前体 mRNA 5 剪接位点。
DOI：
10.1073/pnas.91.8.3363
复制
发表时间：
1994
影响因子：
11.1
通讯作者：
Manley,JL
中科院分区：
文献类型：
--
作者：
Zuo,P;Manley,JL
ASF/SF 2是一种人类蛋白质，以前显示在体外前mRNA剪接中作为所有剪接所必需的必需因子以及作为能够切换5'剪接位点选择的选择性剪接因子发挥作用。为了开始研究蛋白的作用机制，我们研究了纯化的重组ASF/SF 2的RNA结合特性。使用UV交联和凝胶位移分析，我们证明ASF/SF 2的RNA结合区可以以序列特异性方式与RNA相互作用，识别两种不同前体mRNA中的每一种的5'剪接位点。5'剪接位点共有序列中的点突变可使结合减少多达100倍，在竞争测定中观察到最大的影响。这些发现支持ASF/SF 2有助于识别前体mRNA 5'剪接位点的模型。
ASF/SF2 is a human protein previously shown to function in in vitro pre-mRNA splicing as an essential factor necessary for all splices and also as an alternative splicing factor, capable of switching selection of 5' splice sites. To begin to study the protein's mechanism of action, we have investigated the RNA binding properties of purified recombinant ASF/SF2. Using UV crosslinking and gel shift assays, we demonstrate that the RNA binding region of ASF/SF2 can interact with RNA in a sequence-specific manner, recognizing the 5' splice site in each of two different pre-mRNAs. Point mutations in the 5' splice site consensus can reduce binding by as much as a factor of 100, with the largest effects observed in competition assays. These findings support a model in which ASF/SF2 aids in the recognition of pre-mRNA 5' splice sites.