Identification of peptide inhibitors of pre-mRNA splicing derived from the essential interaction domains of CDC5L and PLRG1
Identification of peptide inhibitors of pre-mRNA splicing derived from the essential interaction domains of CDC5L and PLRG1
复制标题
DOI：
10.1093/nar/gkg817
复制
发表时间：
2003-11-01
影响因子：
14.9
通讯作者：
Lamond, AI
中科院分区：
文献类型：
--
作者：
Ajuh, P;Lamond, AI
CDC 5L和PLRG 1都是剪接体蛋白，在物种间高度保守。它们都已被证明是亚剪接体蛋白复合物的一部分，其对于酵母和人类中的前mRNA剪接是必需的。CDC 5L和PLRG 1在体外直接相互作用。这种相互作用由PLRG 1中的WD 40区域和CDC 5L的C-末端结构域介导。为了确定这种相互作用是否对剪接机制很重要，我们设计了与两种蛋白质相互作用结构域中高度保守序列相对应的肽。这些肽在体外剪接实验中用作内源性蛋白质中同源序列的竞争者。发现源自两种蛋白质的结合结构域的某些肽抑制体外剪接。这种剪接抑制可以通过将肽与已在大肠杆菌中表达的相应伴侣蛋白预孵育来防止。本研究的结果表明，CDC 5L和PLRG 1之间的相互作用是必不可少的前体mRNA剪接，并进一步证明，小肽可以用作有效的剪接抑制剂。
CDC5L and PLRG1 are both spliceosomal proteins that are highly conserved across species. They have both been shown to be part of sub- spliceosomal protein complexes that are essential for pre-mRNA splicing in yeast and humans. CDC5L and PLRG1 interact directly in vitro. This interaction is mediated by WD40 regions in PLRG1 and the C-terminal domain of CDC5L. In order to determine whether this interaction is important for the splicing mechanism, we have designed peptides corresponding to highly conserved sequences in the interaction domains of both proteins. These peptides were used in in vitro splicing experiments as competitors to the cognate sequences in the endogenous proteins. Certain peptides derived from the binding domains of both proteins were found to inhibit in vitro splicing. This splicing inhibition could be prevented by preincubating the peptides with the corresponding partner protein that had been expressed in Escherichia coli. The results from this study indicate that the interaction between CDC5L and PLRG1 is essential for pre-mRNA splicing and further demonstrate that small peptides can be used as effective splicing inhibitors.