ISOLATION OF TFC1, A GENE ENCODING ONE OF 2 DNA-BINDING SUBUNITS OF YEAST TRANSCRIPTION FACTOR-TAU (TFIIIC)

被引:56
作者
SWANSON, RN
CONESA, C
LEFEBVRE, O
CARLES, C
RUET, A
QUEMENEUR, E
GAGNON, J
SENTENAC, A
机构
[1] CENS,INGN PROT LAB,F-91191 GIF SUR YVETTE,FRANCE
[2] CEN,UNITE CNRS,SERV BIOL STRUCT,F-38041 GRENOBLE,FRANCE
关键词
RNA POLYMERASE-C; TRANSFER RNA; 5S RNA; CLONING;
D O I
10.1073/pnas.88.11.4887
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
Transcription factor TFIIIC mediates tRNA and 5S RNA gene activation by binding to intragenic promoter elements. The factor from Saccharomyces cerevisiae, also called tau, is a large, multisubunit protein (550-650 kDa) containing two polypeptides that interact directly with DNA encoding tRNA (tDNA). We have obtained peptide sequences from the 95-kDa DNA-binding subunit (tau-95) and cloned the corresponding gene, called TFC1. The gene encodes a polypeptide of calculated M(r) 73,500. However, when TFC1 was transcribed and translated in vitro, the gene product comigrated with tau-95 in SDS/polyacrylamide gels. A fusion protein expressed in bacteria was able to prevent the binding of anti-tau-95 antibodies to tau-tDNA complexes. The TFC1 gene is present in single copy on yeast chromosome II and is essential for growth. Spores containing a disrupted gene germinate but only proceed through a few cell divisions before ceasing to grow. The TFC1-encoded protein contains a potential helix-turn-helix structure and an acidic carboxyl-terminal domain, a feature characteristic of some DNA-binding proteins and transcriptional regulators.
引用
收藏
页码:4887 / 4891
页数:5
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