Journal of Biosciences

, Volume 36, Issue 1, pp 43–54 | Cite as

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

  • Olga N KorolevaEmail author
  • Stephen JW Busby
  • Valeriy L Drutsa


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.


Non-conserved region RNA polymerase σ subunit site-specific mutagenesis transcription 



Intein-Mediated Purification with an Affinity Chitin-binding Tag


non-conserved region


RNA polymerase


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Copyright information

© Indian Academy of Sciences 2011

Authors and Affiliations

  • Olga N Koroleva
    • 1
    Email author
  • Stephen JW Busby
    • 2
  • Valeriy L Drutsa
    • 3
  1. 1.Chemical FacultyLomonosov Moscow State UniversityMoscowRussia
  2. 2.School of BiosciencesThe University of BirminghamBirminghamUK
  3. 3.Belozersky Institute of Physico-Chemical BiologyLomonosov Moscow State UniversityMoscowRussia

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