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J. Biol. Chem., Vol. 278, Issue 50, 50588-50595, December 12, 2003

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Signal-dependent Requirement for the Co-activator Protein RcsA in Transcription of the RcsB-regulated ugd Gene^*

Chakib Mouslim, Tammy Latifi, and Eduardo A. Groisman

From the Department of Molecular Microbiology, Howard Hughes Medical Institute, Washington University School of Medicine, St. Louis, Missouri 63110

The RcsC/YojN/RcsB phosphorelay system controls gene expression in response to a variety of signals, including changes in temperature, osmolarity, and overproduction of membrane proteins. Transcription of certain RcsB-activated genes, such as the capsule synthesis cps operon, requires the co-activator protein RcsA, whereas expression of other RcsB-activated genes is RcsA-independent. We have established previously that a tolB mutation induces transcription of the Salmonella UDP-glucose dehydrogenase ugd gene in an RcsA- and RcsB-dependent manner. This induction is independent of the two-component systems PhoP/PhoQ and PmrA/PmrB, which are required for ugd expression in response to low Mg²⁺. We now report that the RcsC/YojN/RcsB system is activated in a pmrA mutant experiencing Fe³⁺ and low Mg²⁺, resulting in expression of both cps and ugd genes. However, whereas cps transcription remained RcsA-dependent, ugd transcription became RcsA-independent but dependent on the PhoP protein. S1 mapping experiments demonstrated that RcsA-dependent and -independent transcription of the ugd gene use the same promoter. DNase footprinting analysis identified a PhoP-binding site in the ugd promoter. Yet, PhoP-mediated ugd transcription required either the RcsC/YojN/RcsB or the PmrA/PmrB systems.

Received for publication, August 26, 2003 , and in revised form, September 22, 2003.

^* This work was supported by National Institutes of Health Grants AI42336 and AI49561 (to E. A. 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.

Investigator of the Howard Hughes Medical Institute. To whom correspondence should be addressed: Dept. of Molecular Microbiology, Howard Hughes Medical Institute, Washington Univ. School of Medicine, Campus Box 8230, 660 S. Euclid, St. Louis, MO 63110. Tel.: 314-362-3692; Fax: 314-747-8228; E-mail: groisman{at}borcim.wustl.edu.

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