Prp22, a DExH-box RNA helicase, plays two distinct roles in yeast pre-mRNA splicing
Prp22, a DExH-box RNA helicase, plays two distinct roles in yeast pre-mRNA splicing
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DOI：
10.1093/emboj/17.7.2086
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发表时间：
1998-04-01
期刊：
影响因子：
11.4
通讯作者：
Gross, CH
中科院分区：
文献类型：
--
作者：
Schwer, B;Gross, CH
为了评估Prp 22在酵母前体mRNA剪接中的作用，我们纯化了130 kDa的Prp 22蛋白，并开发了体外耗竭/重建测定。我们表明，Prp 22是所需的肌动蛋白前mRNA剪接的第二步。Prp 22可以作用于预组装的剪接体，这些剪接体在步骤1后以ATP-独立的方式被捕获。在步骤2中对Prp 22的需求取决于分支点和3'剪接位点之间的距离，这表明Prp 22在剪接位点选择中的作用是以前未认识到的。我们的特点的生化活动Prp 22，DExH盒蛋白质家族的成员，我们表明，纯化的重组Prp 22蛋白是一个RNA依赖的ATP酶和ATP依赖的RNA解旋酶。Prp 22利用ATP水解的能量来实现mRNA从剪接体的释放。因此，Prp 22在酵母前体mRNA剪接中具有两种不同的功能：在第二催化步骤中的ATP-独立作用和在剪接体的拆解中的ATP-需要功能。
In order to assess the role of Prp22 in yeast pre-mRNA splicing, we have purified the 130 kDa Prp22 protein and developed an in vitro depletion/reconstitution assay. We show that Prp22 is required for the second step of actin pre-mRNA splicing. Prp22 can act on preassembled spliceosomes that are arrested after step 1 in an ATP-independent fashion. The requirement for Prp22 during step 2 depends on the distance between the branchpoint and the 3' splice site, suggesting a previously unrecognized role for Prp22 in splice site selection. We characterize the biochemical activities of Prp22, a member of the DExH-box family of proteins, and we show that purified recombinant Prp22 protein is an RNA-dependent ATPase and an ATP-dependent RNA helicase. Prp22 uses the energy of ATP hydrolysis to effect the release of mRNA from the spliceosome. Thus, Prp22 has two distinct functions in yeast pre-mRNA splicing: an ATP-independent role during the second catalytic step and an ATP-requiring function in disassembly of the spliceosome.