Nucleic Acids Research Advance Access originally published online on March 27, 2007
Nucleic Acids Research 2007 35(7):2311-2320; doi:10.1093/nar/gkm123
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Nucleic Acids Research, 2007, Vol. 35, No. 7 2311-2320
© 2007 The Author(s)
This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
Molecular Biology |
The C-terminal domain of the Escherichia coli RNA polymerase 
subunit plays a role in the CI-dependent activation of the bacteriophage 
pM promoter
1Department of Molecular Biology, University of Gdansk, Kladki 24, 80-822 Gdansk, Poland, 2School of Biosciences, University of Birmingham, Edgbaston, Birmingham B15 2TT, UK and 3School of Medicine and Biomedical Sciences, University of Sheffield, Beech Hill Road, Sheffield S10 2RX, UK
*To whom correspondence should be addressed. Tel: +44 114 271 2834; Fax: +44 114 271 3892; Email: m.s.thomas{at}shef.ac.uk
Received January 18, 2007. Revised February 14, 2007. Accepted February 14, 2007.
The bacteriophage 
pM promoter is required for maintenance of the prophage in Escherichia coli, as it facilitates transcription of the cI gene, encoding the repressor (CI). CI levels are maintained through a transcriptional feedback mechanism whereby CI can serve as an activator or a repressor of pM. CI activates pM through cooperative binding to the OR1 and OR2 sites within the OR operator, with the OR2-bound CI dimer making contact with domain 4 of the RNA polymerase 
subunit (4). Here we demonstrate that the 261 and 287 determinants of the C-terminal domain of the RNA polymerase 
subunit (CTD), as well as the DNA-binding determinant, are important for CI-dependent activation of pM. We also show that the location of CTD at the pM promoter changes in the presence of CI. Thus, in the absence of CI, one CTD is located on the DNA at position –44 relative to the transcription start site, whereas in the presence of CI, CTD is located at position –54, between the CI-binding sites at OR1 and OR2. These results suggest that contacts between CI and both CTD and are required for efficient CI-dependent activation of pM.
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