Construction and functional analyses of a comprehensive Sigma(54)   site-directed mutant library using alanine-cysteine mutagenesis

doi:10.1093/nar/gkp419

CORC > 北京大学 > 生命科学学院

	Construction and functional analyses of a comprehensive Sigma(54) site-directed mutant library using alanine-cysteine mutagenesis
	Xiao, Yan ; Wigneshweraraj, Siva R. ; Weinzierl, Robert ; Wang, Yi-Ping ; Buck, Martin
刊名	核酸研究
	2009
关键词	COLI RNA-POLYMERASE ESCHERICHIA-COLI DNA-BINDING PROMOTER DNA TRANSCRIPTIONAL ACTIVATION ENHANCER RESPONSIVENESS TERMINAL SEQUENCES FACTOR SIGMA-54 CROSS-LINKING IN-VIVO
DOI	10.1093/nar/gkp419
英文摘要	The Sigma(54) factor associates with core RNA polymerase (RNAP) to form a holoenzyme that is unable to initiate transcription unless acted on by an activator protein. Sigma(54) is closely involved in many steps of activator-dependent transcription, such as core RNAP binding, promoter recognition, activator interaction and open complex formation. To systematically define Sigma(54) residues that contribute to each of these functions and to generate a resource for site specific protein labeling, a complete mutant library of Sigma(54) was constructed by alanine-cysteine scanning mutagenesis. Amino acid residues from 3 to 476 of Cys(-)Sigma(54) were systematically mutated to alanine and cysteine in groups of two adjacent residues at a time. The influences of each substitution pair upon the functions of Sigma(54) were analyzed in vivo and in vitro and the functions of many residues were revealed for the first time. Increased Sigma(54) isomerization activity seldom corresponded with an increased transcription activity of the holoenzyme, suggesting the steps after Sigma(54) isomerization, likely to be changes in core RNAP structure, are also strictly regulated or rate limiting to open complex formation. A linkage between core RNAP-binding activity and activator responsiveness indicates that the Sigma(54)-core RNAP interface changes upon activation.; Biochemistry & Molecular Biology; SCI(E); PubMed; 3; ARTICLE; 13; 4482-4497; 37
语种	英语
内容类型	期刊论文
源URL	[http://ir.pku.edu.cn/handle/20.500.11897/319989]
专题	生命科学学院
推荐引用方式 GB/T 7714	Xiao, Yan,Wigneshweraraj, Siva R.,Weinzierl, Robert,et al. Construction and functional analyses of a comprehensive Sigma(54) site-directed mutant library using alanine-cysteine mutagenesis[J]. 核酸研究,2009.
APA	Xiao, Yan,Wigneshweraraj, Siva R.,Weinzierl, Robert,Wang, Yi-Ping,&Buck, Martin.(2009).Construction and functional analyses of a comprehensive Sigma(54) site-directed mutant library using alanine-cysteine mutagenesis.核酸研究.
MLA	Xiao, Yan,et al."Construction and functional analyses of a comprehensive Sigma(54) site-directed mutant library using alanine-cysteine mutagenesis".核酸研究 (2009).