Binding of a cell-type-specific RNA splicing factor to its target regulatory sequence.
Binding of a cell-type-specific RNA splicing factor to its target regulatory sequence.
复制标题
细胞类型特异性 RNA 剪接因子与其靶标调控序列的结合。
DOI：
10.1128/mcb.15.4.1953
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发表时间：
1995
影响因子：
5.3
通讯作者：
Roeder,GS
中科院分区：
文献类型：
--
作者：
Nandabalan,K;Roeder,GS
酿酒酵母MER 2基因的转录本在减数分裂过程中有效剪接，但在营养生长过程中不剪接。野生型MER 2转录本的有效拼接需要Mer 1蛋白，该蛋白仅在减数分裂细胞中产生。对MER 25 ′外显子缺失和替换突变的分析表明，该外显子异常大的尺寸在剪接调控中起重要作用。通过对MER 2缺失突变体和杂合基因的分析，确定了MER 2 RNA的MER 1依赖性剪接所必需的顺式作用序列。80碱基的MER 2内含子足以在体内进行Mer1依赖性剪接，但5′外显子中的序列增强剪接效率。Mer1蛋白含有在一些RNA结合蛋白中发现的KH基序，RNA凝胶迁移率变动分析表明Mer1特异性结合MER2RNA。无论是来自内含子的MER 2基因的转录本和转录本组成的内含子都能够结合到MER 1在体外，但没有作为完整的底物的蛋白质的高亲和力。RNA酶T1足迹法表明，Mer1蛋白在5′外显子和内含子的几个位点上与MER2RNA接触。因此，Mer1直接与MER2RNA中的调控元件相互作用并促进剪接。
The transcript of theSaccharomyces cerevisiae MER2gene is spliced efficiently during meiosis but not during vegetative growth. Efficient splicing of the wild-typeMER2transcript requires the Mer1 protein, which is produced only in meiotic cells. Analysis of deletion and substitution mutations in theMER25′ exon demonstrates that the unusually large size of this exon plays an important role in splicing regulation. Thecis-acting sequences essential for Mer1-dependent splicing ofMER2RNA were determined by the analysis ofMER2deletion mutants and hybrid genes. The 80-baseMER2intron is sufficient for Mer1-dependent splicing in vivo, but sequences in the 5′ exon enhance splicing efficiency. The Mer1 protein contains the KH motif found in some RNA-binding proteins, and RNA gel mobility shift assays demonstrate that Mer1 binds specifically toMER2RNA. Both the transcript derived from the intronlessMER2gene and the transcript consisting only of the intron are able to bind to Mer1 in vitro, but neither has as high affinity for the protein as the intact substrate. RNase T1footprinting indicates that the Mer1 protein contactsMER2RNA at several points in the 5′ exon and in the intron. Thus, Mer1 interacts directly with a regulatory element inMER2RNA and promotes splicing.