Prp22, a DExH-box RNA helicase, plays two distinct roles in yeast pre-mRNA splicing

被引:194
作者
Schwer, B [1 ]
Gross, CH
机构
[1] Cornell Univ, Coll Med, Dept Microbiol, New York, NY 10021 USA
[2] Sloan Kettering Inst, Program Mol Biol, New York, NY 10021 USA
关键词
ATPase; DExH-box; Prp22; RNA helicase; spliceosome;
D O I
10.1093/emboj/17.7.2086
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
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.
引用
收藏
页码:2086 / 2094
页数:9
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