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Journal of Bacteriology, February 2004, p. 1165-1174, Vol. 186, No. 4
0021-9193/04/$08.00+0     DOI: 10.1128/JB.186.4.1165-1174.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

clpB, a Novel Member of the Listeria monocytogenes CtsR Regulon, Is Involved in Virulence but Not in General Stress Tolerance

Arnaud Chastanet,1,{dagger} Isabelle Derre,1,{dagger} Shamila Nair,2 and Tarek Msadek1*

Unité de Biochimie Microbienne, Institut Pasteur, CNRS URA 2172, 75724 Paris Cedex 15,1 INSERM U411, Faculté de Médecine Necker, 156, 75730 Paris Cedex 15, France2

Received 2 July 2003/ Accepted 23 October 2003

Clp-HSP100 ATPases are a widespread family of ubiquitous proteins that occur in both prokaryotes and eukaryotes and play important roles in the folding of newly synthesized proteins and refolding of aggregated proteins. They have also been shown to participate in the virulence of several pathogens, including Listeria monocytogenes. Here, we describe a member of the Clp-HSP100 family of L. monocytogenes that harbors all the characteristics of the ClpB subclass, which is absent in the closely related gram-positive model organism, Bacillus subtilis. Transcriptional analysis of clpB revealed a heat shock-inducible {sigma}A-type promoter. Potential binding sites for the CtsR regulator of stress response were identified in the promoter region. In vivo and in vitro approaches were used to show that expression of clpB is repressed by CtsR, a finding indicating that clpB is a novel member of the L. monocytogenes CtsR regulon. We showed that ClpB is involved in the pathogenicity of L. monocytogenes since the {Delta}clpB mutant is significantly affected by virulence in a murine model of infection; we also demonstrate that this effect is apparently not due to a defect in general stress resistance. Indeed, ClpB is not involved in tolerance to heat, salt, detergent, puromycin, or cold stress, even though its synthesis is inducible by heat shock. However, ClpB was shown to play a role in induced thermotolerance, allowing increased resistance of L. monocytogenes to lethal temperatures. This work gives the first example of a clpB gene directly controlled by CtsR and describes the first role for a ClpB protein in induced thermotolerance and virulence in a gram-positive organism.

* Corresponding author. Mailing address: Unité de Biochimie Microbienne, Institut Pasteur, 25 rue du Dr. Roux, 75724 Paris Cedex 15, France. Phone: 33-1-45-68-88-09. Fax: 33-1-45-68-89-38. E-mail: tmsadek{at}pasteur.fr.

{dagger} A.C. and I.D. contributed equally to this report.

Journal of Bacteriology, February 2004, p. 1165-1174, Vol. 186, No. 4
0021-9193/04/$08.00+0     DOI: 10.1128/JB.186.4.1165-1174.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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