果蝇转录起始因子TFIIF小亚基编码cDNA的分子克隆

Molecular cloning of cDNA encoding the small subunit of Drosophila transcription initiation factor TFIIF.

作者信息

Gong D W, Mortin M A, Horikoshi M, Nakatani Y

机构信息

National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD 20892, USA.

出版信息

Nucleic Acids Res. 1995 Jun 11;23(11):1882-6. doi: 10.1093/nar/23.11.1882.

DOI:10.1093/nar/23.11.1882

PMID:7596813

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC306958/

Abstract

Transcription initiation factor TFIIF is a tetramer consisting of two large subunits (TFIIF alpha or RAP74) and two small subunits (TFIIF beta or RAP30). We report here the molecular cloning of a Drosophila cDNA encoding TFIIF beta. The cDNA clone contains an open-reading frame encoding a 277 amino acid polypeptide having a calculated molecular mass of 32,107 Da. Comparison of the deduced amino acid sequence with the corresponding sequences from vertebrates showed only 50% identity, with four insertion/deletion points. For transcription activity in a TFIIF-depleted Drosophila nuclear extract, both TFIIF alpha and TFIIF beta are essential. Moreover, Drosophila TFIIF beta interacts with both Drosophila and human TFIIF alpha in vitro. Thus we conclude that isolated cDNA encodes bona fide TFIIF beta. The structural domains of TFIIF beta and its sequence similarity to bacterial delta factors are discussed.

摘要

转录起始因子TFIIF是一种四聚体，由两个大亚基（TFIIFα或RAP74）和两个小亚基（TFIIFβ或RAP30）组成。我们在此报告编码TFIIFβ的果蝇cDNA的分子克隆。该cDNA克隆包含一个开放阅读框，编码一个277个氨基酸的多肽，计算分子量为32,107道尔顿。将推导的氨基酸序列与脊椎动物的相应序列进行比较，发现只有50%的同一性，有四个插入/缺失位点。对于在缺乏TFIIF的果蝇核提取物中的转录活性，TFIIFα和TFIIFβ都是必不可少的。此外，果蝇TFIIFβ在体外与果蝇和人类TFIIFα都相互作用。因此我们得出结论，分离的cDNA编码真正的TFIIFβ。本文还讨论了TFIIFβ的结构域及其与细菌δ因子的序列相似性。