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Journal of Bacteriology, December 2001, p. 7295-7307, Vol. 183, No. 24
0021-9193/01/$04.00+0   DOI: 10.1128/JB.183.24.7295-7307.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Regulation of Streptococcus pneumoniae clp Genes and Their Role in Competence Development and Stress Survival

Arnaud Chastanet,1 Marc Prudhomme,2 Jean-Pierre Claverys,2 and Tarek Msadek1,*

Unité de Biochimie Microbienne, Institut Pasteur, URA 2172 du Centre National de la Recherche Scientifique, 75724 Paris Cedex 15,1 and Laboratoire de Microbiologie et de Génétique Moléculaire, UMR5100 du Centre National de la Recherche Scientifique-Université Paul Sabatier, 31062 Toulouse Cedex,2 France

Received 2 July 2001/Accepted 19 September 2001

In vitro mariner transposon mutagenesis of Streptococcus pneumoniae chromosomal DNA was used to isolate regulatory mutants affecting expression of the comCDE operon, encoding the peptide quorum-sensing two-component signal transduction system controlling competence development. A transposon insertion leading to increased comC expression was found to lie directly upstream from the S. pneumoniae clpP gene, encoding the proteolytic subunit of the Clp ATP-dependent protease, whose expression in Bacillus subtilis is controlled by the CtsR repressor. In order to examine clp gene regulation in S. pneumoniae, a detailed analysis of the complete genome sequence was performed, indicating that there are five likely CtsR-binding sites located upstream from the clpE, clpP, and clpL genes and the ctsR-clpC and groESL operons. The S. pneumoniae ctsR gene was cloned under the control of an inducible promoter and used to demonstrate regulation of the S. pneumoniae clpP and clpE genes and the clpC and groESL operons by using B. subtilis as a heterologous host. The CtsR protein of S. pneumoniae was purified and shown to bind specifically to the clpP, clpC, clpE, and groESL regulatory regions. S. pneumoniae Delta ctsR, Delta clpP, Delta clpC, and Delta clpE mutants were constructed by gene deletion/replacement. ClpP was shown to act as a negative regulator, preventing competence gene expression under inappropriate conditions. Phenotypic analyses also indicated that ClpP and ClpE are both required for thermotolerance. Contrary to a previous report, we found that ClpC does not play a major role in competence development, autolysis, pneumolysin production, or growth at high temperature of S. pneumoniae.

* 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.

Journal of Bacteriology, December 2001, p. 7295-7307, Vol. 183, No. 24
0021-9193/01/$04.00+0   DOI: 10.1128/JB.183.24.7295-7307.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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