Effects of substitutions at position 180 in the Escherichia coli RNA polymerase 𝛼70 subunit

Journal of biosciences

Stephen Jw Busby³  Olga N Koroleva1¹  Valeriy L Drutsa² 

[1] Chemical Faculty, Lomonosov Moscow State University, 119992 Moscow, Russia$$;Belozersky Institute of Physico-Chemical Biology, Lomonosov Moscow State University, 119992 Moscow, Russia$$;School of Biosciences, The University of Birmingham, Edgbaston, Birmingham B15 2TT, UK$$

关键词: Non-conserved region;    RNA polymerase;    𝛼 subunit;    site-specific mutagenesis;    transcription;   

DOI  :  

来源: Indian Academy of Sciences

【 摘 要 】

In order to investigate the role of His180 residue, located in the non-conserved region of the 𝜎70 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 𝜎70 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.

【 授权许可】

Unknown