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Published online 2 January 2004

Nucleic Acids Research, 2004, Vol. 32, No. 1 45-53
© 2004 Oxford University Press

Interactions between the 2.4 and 4.2 regions of {sigma}S, the stress-specific {sigma} factor of Escherichia coli, and the –10 and –35 promoter elements

Claire Checroun, Patricia Bordes, Olivier Leroy1, Annie Kolb1 and Claude Gutierrez*

Laboratoire de Microbiologie et Génétique Moléculaire, UMR5100 CNRS–Université Toulouse III, 118, Route de Narbonne, 31062, Toulouse Cedex, France and 1 Unité des Régulations Transcriptionnelles, Institut Pasteur, 25 rue du Dr Roux, 75724 Paris cedex 15, France

*To whom correspondence should be addressed. Tel: +33 5 61 33 58 72; Fax: +33 5 61 33 58 86; Email: clg{at}ibcg.biotoul.fr
Present address:
Patricia Bordes, Department of Biological Sciences, Sir Alexander Fleming Building, Imperial College London, South Kensington Campus, London SW7 2AZ, UK

The {sigma}s subunit of Escherichia coli RNA polymerase holoenzyme (E{sigma}S) is a key factor of gene expression upon entry into stationary phase and in stressful conditions. The selectivity of promoter recognition by E{sigma}S and the housekeeping E{sigma}70 is as yet not clearly understood. We used a genetic approach to investigate the interaction of {sigma}S with its target promoters. Starting with down-promoter variants of a {sigma}S promoter target, osmEp, altered in the –10 or –35 elements, we isolated mutant forms of {sigma}S suppressing the promoter defects. The activity of these suppressors on variants of osmEp and ficp, another target of {sigma}S, indicated that {sigma}S is able to interact with the same key features within a promoter sequence as {sigma}70. Indeed, (i) {sigma}S can recognize the –35 element of some but not all its target promoters, through interactions with its 4.2 region; and (ii) amino acids within the 2.4 region participate in the recognition of the –10 element. More specifically, residues Q152 and E155 contribute to the strong preference of {sigma}S for a C in position –13 and residue R299 can interact with the –31 nucleotide in the –35 element of the target promoters.


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