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Journal of Bacteriology, June 2006, p. 3983-3994, Vol. 188, No. 11
0021-9193/06/$08.00+0     doi:10.1128/JB.00033-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Crl Activates Transcription Initiation of RpoS-Regulated Genes Involved in the Multicellular Behavior of Salmonella enterica Serovar Typhimurium

Véronique Robbe-Saule,1 Valentin Jaumouillé,1,{dagger} Marie-Christine Prévost,2 Stéphanie Guadagnini,2 Christelle Talhouarne,1 Hayette Mathout,1 Annie Kolb,1 and Françoise Norel1*

Unité des Régulations Transcriptionnelles, URA-CNRS 2172,1 Plateforme de Microscopie Electronique, Institut Pasteur, 25 rue du Docteur Roux, 75724 Paris Cedex 15, France2

Received 10 January 2006/ Accepted 15 March 2006

In Salmonella enterica serovar Typhimurium, the stationary-phase sigma factor {sigma}S (RpoS) is required for virulence, stress resistance, biofilm formation, and development of the rdar morphotype. This morphotype is a multicellular behavior characterized by expression of the adhesive extracellular matrix components cellulose and curli fimbriae. The Crl protein of Escherichia coli interacts with {sigma}S and activates expression of {sigma}S-regulated genes, such as the csgBAC operon encoding the subunit of the curli proteins, by an unknown mechanism. Here, we showed using in vivo and in vitro experiments that the Crl protein of Salmonella serovar Typhimurium is required for development of a typical rdar morphotype and for maximal expression of the csgD, csgB, adrA, and bcsA genes, which are involved in curli and cellulose biosynthesis. In vitro transcription assays and potassium permanganate reactivity experiments with purified His6-Crl showed that Crl directly activated {sigma}S-dependent transcription initiation at the csgD and adrA promoters. We observed no effect of Crl on {sigma}70-dependent transcription. Crl protein levels increased during the late exponential and stationary growth phases in Luria-Beratani medium without NaCl at 28°C. We obtained complementation of the crl mutation by increasing {sigma}S levels. This suggests that Crl has a major physiological impact at low concentrations of {sigma}S.

* Corresponding author. Mailing address: Institut Pasteur, Unité des Régulations Transcriptionnelles, URA-CNRS 2172, 25 rue du Docteur Roux, 75724 Paris Cedex 15, France. Phone: 33 140613122. Fax: 33 145688960. E-mail: francoise.norel{at}pasteur.fr.

{dagger} Present address: Unité de Pathogénie Microbienne Institut Pasteur, 28 rue du Docteur Roux, 75724 Paris Cedex 15, France.

Journal of Bacteriology, June 2006, p. 3983-3994, Vol. 188, No. 11
0021-9193/06/$08.00+0     doi:10.1128/JB.00033-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



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