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Journal of Bacteriology, January 2004, p. 179-191, Vol. 186, No. 1
0021-9193/04/$08.00+0     DOI: 10.1128/JB.186.1.179-191.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Fine-Tuning in Regulation of Clp Protein Content in Bacillus subtilis{dagger}

Ulf Gerth,1* Janine Kirstein,1 Jörg Mostertz,1 Torsten Waldminghaus,2 Marcus Miethke,1 Holger Kock,1 and Michael Hecker1

Institut für Mikrobiologie, Ernst-Moritz-Arndt-Universität, D-17487 Greifswald,1 Abteilung Immunologie, Max-Planck-Institut für Infektionsbiologie, D-10117 Berlin, Germany2

Received 7 July 2003/ Accepted 3 October 2003

Clp-controlled proteolysis in Bacillus subtilis seems to play a substantial role, particularly under stress conditions. Calibrated Western blot analyses were used to estimate the approximate numbers of heat-inducible Clp molecules within a single cell. According to these numbers, the different Clp ATPases do not seem to compete for the proteolytic subunit ClpP. Coimmunoprecipitation experiments revealed the predicted specific ClpX-ClpP, ClpC-ClpP, and ClpE-ClpP interactions. ClpE and ClpX are rapidly degraded in wild-type cells during permanent heat stress but remained almost stable in a clpP mutant, suggesting ClpP-dependent degradation. In particular, ClpCP appeared to be involved in the degradation of the short-lived ClpE ATPase, indicating a negative "autoregulatory" circuit for this particular Clp ATPase at the posttranslational level. Analysis of the half-life of stress-inducible clp mRNAs during exponential growth and heat shock revealed precise regulation of the synthesis of each Clp protein at the posttranscriptional level as well to meet the needs of B. subtilis.

* Corresponding author. Mailing address: Ernst-Moritz-Arndt-Universität, Institut für Mikrobiologie, F.-L.-Jahn-Str. 15, D-17487 Greifswald, Germany. Phone: 49-3834-864216. Fax: 49-3834-864202. E-mail: Ulf.Gerth{at}biologie.uni-greifswald.de.

{dagger} This article is dedicated to Sierd Bron (University of Groningen, Groningen, The Netherlands) for his great contribution to Bacillus subtilis genetics.

Journal of Bacteriology, January 2004, p. 179-191, Vol. 186, No. 1
0021-9193/04/$08.00+0     DOI: 10.1128/JB.186.1.179-191.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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