5'剪接位点处的RNA开关需要ATP和DEAD盒蛋白Prp28p。

An RNA switch at the 5' splice site requires ATP and the DEAD box protein Prp28p.

作者信息

Staley J P, Guthrie C

机构信息

Department of Biochemistry and Biophysics, University of California, San Francisco 94143, USA.

出版信息

Mol Cell. 1999 Jan;3(1):55-64. doi: 10.1016/s1097-2765(00)80174-4.

DOI:10.1016/s1097-2765(00)80174-4

PMID:10024879

Abstract

Pre-mRNA splicing requires dramatic RNA rearrangements hypothesized to be catalyzed by ATP-dependent RNA unwindases of the DExD/H box family. In a rearrangement critical for the fidelity of 5' splice site recognition, a base-pairing interaction between the 5' splice site and U1 snRNA must be switched for a mutually exclusive interaction between the 5' splice site and U6 snRNA. By lengthening the U1:5' splice site duplex, we impeded this switch in a temperature-dependent manner and prevented formation of the spliceosome's catalytic core. Using genetics, we identified the DExD/H box protein Prp28p as a potential mediator of the switch. In vitro, the switch requires both Prp28p and ATP. We propose that Prp28p directs isomerization of RNA at the 5' splice site and promotes fidelity in splicing.

摘要

前体mRNA剪接需要显著的RNA重排，据推测这是由DExD/H盒家族的ATP依赖性RNA解旋酶催化的。在对5'剪接位点识别保真度至关重要的重排过程中，5'剪接位点与U1 snRNA之间的碱基配对相互作用必须转换为5'剪接位点与U6 snRNA之间相互排斥的相互作用。通过延长U1:5'剪接位点双链体，我们以温度依赖的方式阻碍了这种转换，并阻止了剪接体催化核心的形成。利用遗传学方法，我们鉴定出DExD/H盒蛋白Prp28p是这种转换的潜在介导因子。在体外，这种转换既需要Prp28p也需要ATP。我们提出，Prp28p指导5'剪接位点处RNA的异构化，并促进剪接的保真度。