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RNA聚合酶II的Rpb7亚基与一种参与转录本加工的RNA结合蛋白相互作用。

Rpb7 subunit of RNA polymerase II interacts with an RNA-binding protein involved in processing of transcripts.

作者信息

Mitsuzawa Hiroshi, Kanda Emi, Ishihama Akira

机构信息

Department of Molecular Genetics, National Institute of Genetics, Mishima, Shizuoka 411-8540, Japan.

出版信息

Nucleic Acids Res. 2003 Aug 15;31(16):4696-701. doi: 10.1093/nar/gkg688.

Abstract

Rpb4-Rpb7, a dissociable subcomplex of RNA polymerase II (pol II), is required for transcription initiation. To understand the role of Rpb7 in transcription initiation or other processes in transcription, we carried out a two-hybrid screen for proteins that interact with Rpb7 of the fission yeast Schizosaccharomyces pombe. The screen identified the S.pombe homolog of the Saccharomyces cerevisiae Nrd1, an RNA-binding protein implicated in 3' end formation of small nucleolar and small nuclear RNAs transcribed by pol II. The S.pombe protein, named Seb1 for seven binding, was essential for cell viability, and bound directly to Rpb7 in vitro. Saccharomyces cerevisiae Rpb7 also interacted with Nrd1, indicating that the interaction is conserved in evolution. Glu166 and/or Asp167 of S.pombe Rpb7, residues near the C-terminus of the 172 amino acid protein, were found to be important for its interaction with Seb1. Our results suggest that Rpb7 may function to anchor a processing factor to the pol II apparatus, thereby coupling RNA processing to transcription. The role for Rpb7 is consistent with its location in the pol II complex determined by recent structural studies.

摘要

Rpb4-Rpb7是RNA聚合酶II(pol II)的一个可解离亚复合物,是转录起始所必需的。为了了解Rpb7在转录起始或转录的其他过程中的作用,我们对与粟酒裂殖酵母的Rpb7相互作用的蛋白质进行了双杂交筛选。该筛选鉴定出了酿酒酵母Nrd1的粟酒裂殖酵母同源物,Nrd1是一种与pol II转录的小核仁RNA和小核RNA的3'末端形成有关的RNA结合蛋白。粟酒裂殖酵母蛋白因与七种蛋白结合而被命名为Seb1,对细胞活力至关重要,并且在体外直接与Rpb7结合。酿酒酵母Rpb7也与Nrd1相互作用,表明这种相互作用在进化中是保守的。发现粟酒裂殖酵母Rpb7的Glu166和/或Asp167(位于172个氨基酸蛋白的C末端附近的残基)对其与Seb1的相互作用很重要。我们的结果表明,Rpb7可能起到将一个加工因子锚定到pol II装置上的作用,从而将RNA加工与转录偶联起来。Rpb7的作用与其在最近的结构研究中确定的在pol II复合物中的位置一致。

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