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J. Biol. Chem., Vol. 282, Issue 46, 33326-33335, November 16, 2007

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SlyA and H-NS Regulate Transcription of the Escherichia coli K5 Capsule Gene Cluster, and Expression of slyA in Escherichia coli Is Temperature-dependent, Positively Autoregulated, and Independent of H-NS^*

David Corbett, Hayley J. Bennett, Hamdia Askar¹, Jeffrey Green^¶, and Ian S. Roberts²

From the Faculty of Life Sciences, University of Manchester, Smith Bldg., Oxford Rd., Manchester M13 9PT, United Kingdom, the Department of Medical Microbiology and Immunology, Faculty of Medicine, Mansoura University, Egypt, and the ^¶Department of Molecular Biology and Biotechnology, University of Sheffield, Sheffield S10 2TN, United Kingdom

In this paper, we present the first evidence of a role for the transcriptional regulator SlyA in the regulation of transcription of the Escherichia coli K5 capsule gene cluster and demonstrate, using a combination of reporter gene fusions, DNase I footprinting, and electrophoretic mobility shift assays, the dependence of transcription on the functional interplay between H-NS and SlyA. Both SlyA and H-NS bind to multiple overlapping sites within the promoter in vitro, but their binding is not mutually exclusive, resulting in a remodeled nucleoprotein complex. In addition, we show that expression of the E. coli slyA gene is temperature-regulated, positively autoregulated, and independent of H-NS.

Received for publication, April 25, 2007 , and in revised form, August 30, 2007.

^* This work was supported by a Biotechnology and Biological Sciences Research Council (BBSRC) studentship (to D. C.), by a BBSRC project grant (to I. S. R.), and a Wellcome Trust project grant (to J. G.). The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

The on-line version of this article (available at http://www.jbc.org) contains supplemental Fig. S1.

¹ Supported by an Egyptian Government scholarship.

² To whom correspondence should be addressed. Tel.: 161-275-5601; Fax: 161-275-5656; E-mail: i.s.roberts{at}manchester.ac.uk.

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