In Vivo and In Vitro Effects of (p)ppGpp on the ς 54 Promoter Pu of the TOL Plasmid of Pseudomonas putida
10.1128/jb.182.17.4711-4718.2000
Crossref journal-article
American Society for Microbiology
Journal of Bacteriology (235)
Abstract

ABSTRACT The connection between the physiological control of the ς 54 -dependent Pu promoter of the TOL plasmid pWW0 of Pseudomonas putida and the stringent response mediated by the alarmone (p)ppGpp has been examined in vivo an in vitro. To this end, the key regulatory elements of the system were faithfully reproduced in an Escherichia coli strain and assayed as lacZ fusions in various genetic backgrounds lacking (p)ppGpp or overexpressing relA . Neither the responsiveness of Pu to 3-methyl benzylalcohol mediated by its cognate activator XylR nor the down-regulation of the promoter by rapid growth were affected in relA/spoT strains to an extent which could account for the known physiological control that governs this promoter. Overexpression of the relA gene [predicted to increase intracellullar (p)ppGpp levels] did, however, cause a significant gain in Pu activity. Since such a gain might be the result of indirect effects, we resorted to an in vitro transcription system to assay directly the effect of ppGpp on the transcriptional machinery. Although we did observe a significant increase in Pu performance through a range of ς 54 -RNAP concentrations, such an increase never exceeded twofold. The difference between these results and the behavior of the related Po promoter of the phenol degradation plasmid pVI150 could be traced to the different promoter sequences, which may dictate the type of metabolic signals recruited for the physiological control of ς 54 -systems.

Bibliography

Carmona, M., Rodríguez, M. J., Martínez-Costa, O., & de Lorenzo, V. (2000). In Vivo and In Vitro Effects of (p)ppGpp on the ς 54 Promoter Pu of the TOL Plasmid of Pseudomonas putida. Journal of Bacteriology, 182(17), 4711–4718.

Authors 4
  1. Manuel Carmona (first)
  2. Maria J. Rodríguez (additional)
  3. Óscar Martínez-Costa (additional)
  4. Víctor de Lorenzo (additional)
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Dates
Type When
Created 23 years, 1 month ago (July 27, 2002, 6:03 a.m.)
Deposited 4 years ago (July 29, 2021, 1:54 p.m.)
Indexed 1 year, 5 months ago (March 13, 2024, 9:10 a.m.)
Issued 24 years, 11 months ago (Sept. 1, 2000)
Published 24 years, 11 months ago (Sept. 1, 2000)
Published Print 24 years, 11 months ago (Sept. 1, 2000)
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@article{Carmona_2000, title={In Vivo and In Vitro Effects of (p)ppGpp on the ς 54 Promoter Pu of the TOL Plasmid of Pseudomonas putida}, volume={182}, ISSN={1098-5530}, url={http://dx.doi.org/10.1128/jb.182.17.4711-4718.2000}, DOI={10.1128/jb.182.17.4711-4718.2000}, number={17}, journal={Journal of Bacteriology}, publisher={American Society for Microbiology}, author={Carmona, Manuel and Rodríguez, Maria J. and Martínez-Costa, Óscar and de Lorenzo, Víctor}, year={2000}, month=sep, pages={4711–4718} }