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Journal of Bacteriology, June 2003, p. 3379-3383, Vol. 185, No. 11
0021-9193/03/$08.00+0     DOI: 10.1128/JB.185.11.3379-3383.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Sigma 54 Levels and Physiological Control of the Pseudomonas putida Pu Promoter

Paola Jurado, Luis A. Fernández, and Víctor de Lorenzo*

Centro Nacional de Biotecnología del Consejo Superior de Investigaciones Científicas, Campus de Cantoblanco, 28049 Madrid, Spain

Received 28 February 2003/ Accepted 10 March 2003

The cellular levels of the alternative sigma factor {sigma}54 of Pseudomonas putida have been examined in a variety of growth stages and culture conditions with a single-chain Fv antibody tailored for detection of scarce proteins. The levels of {sigma}54 were also monitored in P. putida strains with knockout mutations in ptsO or ptsN, known to be required for the C-source control of the {sigma}54-dependent Pu promoter of the TOL plasmid. Our results show that ~80 ± 26 molecules of {sigma}54 exist per cell. Unlike that in relatives of Pseudomonas (e.g., Caulobacter), where fluctuations of {sigma}54 determine adaptation and differentiation when cells face starvation, {sigma}54 in P. putida remains unexpectedly constant at different growth stages, in nitrogen starvation and C-source repression conditions, and in the ptsO and ptsN mutant strains analyzed. The number of {sigma}54 molecules per cell in P. putida is barely above the predicted number of {sigma}54-dependent promoters. These figures impose a framework on the mechanism by which Pu (and other {sigma}54-dependent systems) may become amenable to physiological control.

* Corresponding author. Mailing address: Centro Nacional de Biotecnología CSIC, Campus de Cantoblanco, Madrid 28049, Spain. Phone: 34 91 585 45 36. Fax: 34 91 585 45 06. E-mail: vdlorenzo{at}cnb.uam.es.

Journal of Bacteriology, June 2003, p. 3379-3383, Vol. 185, No. 11
0021-9193/03/$08.00+0     DOI: 10.1128/JB.185.11.3379-3383.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

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  • Velazquez, F., Fernandez, S., de Lorenzo, V. (2006). The Upstream-activating Sequences of the {sigma}54 Promoter Pu of Pseudomonas putida Filter Transcription Readthrough from Upstream Genes. J. Biol. Chem. 281: 11940-11948 [Abstract] [Full Text]  
  • Carmona, M., Fernandez, S., Rodriguez, M. J., de Lorenzo, V. (2005). m-Xylene-Responsive Pu-PnifH Hybrid {sigma}54 Promoters That Overcome Physiological Control in Pseudomonas putida KT2442. J. Bacteriol. 187: 125-134 [Abstract] [Full Text]  


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