Effects of substitutions at position 180 in the Escherichia coli RNA polymerase σ70 subunit

被引：3

作者：

Koroleva, Olga N. ^{[1
]}

Busby, Stephen J. W. ^{[2
]}

Drutsa, Valeriy L. ^{[3
]}

机构：

[1] Moscow MV Lomonosov State Univ, Fac Chem, Moscow 119992, Russia

[2] Univ Birmingham, Sch Biosci, Birmingham B15 2TT, W Midlands, England

[3] Moscow MV Lomonosov State Univ, Belozersky Inst Physicochem Biol, Moscow 119992, Russia

来源：

JOURNAL OF BIOSCIENCES | 2011年 / 36卷 / 01期

关键词：

Non-conserved region; RNA polymerase; sigma subunit; site-specific mutagenesis; transcription; AROMATIC-AMINO-ACIDS; SIGMA-70; SUBUNIT; PROMOTER; TRANSCRIPTION; SEQUENCE; PROTEIN; OLIGONUCLEOTIDES; RECOGNITION; MUTAGENESIS; MUTATIONS;

D O I：

10.1007/s12038-011-9007-3

中图分类号：

Q [生物科学];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

In order to investigate the role of His180 residue, located in the non-conserved region of the sigma(70) subunit of Escherichia coli RNA polymerase, two mutant variants of the protein with substitutions for either alanine or glutamic acid were constructed and purified using the IMPACT system. The ability of mutant sigma(70) subunits to interact with core RNA polymerase was investigated using native gel-electrophoresis. The properties of the corresponding reconstituted holoenzymes, as provided by gel shift analysis of their complexes with single- and double-stranded promoter-like DNA and by in vitro transcription experiments, allowed one to deduce that His180 influences several steps of transcription initiation, including core binding, promoter DNA recognition and open complex formation.

引用

页码：43 / 54

页数：12

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