Integration of Signals through Crc and PtsN in Catabolite Repression of Pseudomonas putida TOL Plasmid pWW0

Toluene degradation in Pseudomonas putida KT2440 pWW0 plasmid is subjected to catabolite repression. Pu and P[subscript S1] promoters of the pWW0 TOL plasmid are down-regulated in vivo during exponential growth in rich medium. In cells growing on minimal medium, yeast extract (YE) addition mimics ex...

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Published in:	Applied and Environmental Microbiology 2005-08, Vol.71 (8), p.4191-4198
Main Authors:	Aranda-Olmedo, Isabel, Ramos, Juan L, Marqués, Silvia
Format:	Article
Language:	eng
Subjects:	Bacteria Bacterial Proteins - genetics Bacterial Proteins - metabolism Bacteriology Biodegradation Biodegradation, Environmental Biological and medical sciences catabolite repression crc gene Fundamental and applied biological sciences. Psychology gene down-regulation gene expression regulation Gene Expression Regulation, Bacterial genes genetic techniques and protocols Genetics Genetics and Molecular Biology Integration macroarray technology messenger RNA metabolism Microbiology mutants Mutation Oligonucleotide Array Sequence Analysis - methods Operon Phosphoenolpyruvate Sugar Phosphotransferase System - genetics Phosphoenolpyruvate Sugar Phosphotransferase System - metabolism Plasmids promoter regions Promoter Regions, Genetic Pseudomonas putida Pseudomonas putida - genetics Pseudomonas putida - growth & development Pseudomonas putida - metabolism Repressor Proteins - genetics Repressor Proteins - metabolism Signal Transduction toluene Toluene - metabolism transcription (genetics) transcription factors
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Summary:	Toluene degradation in Pseudomonas putida KT2440 pWW0 plasmid is subjected to catabolite repression. Pu and P[subscript S1] promoters of the pWW0 TOL plasmid are down-regulated in vivo during exponential growth in rich medium. In cells growing on minimal medium, yeast extract (YE) addition mimics exponential-phase rich medium repression of these promoters. We have constructed and tested mutants in a series of global regulators described in PSEUDOMONAS: We describe that a mutant in crc (catabolite repression control) partially relieves YE repression. Macroarray experiments show that crc transcription is strongly increased in the presence of YE, inversely correlated with TOL pathway expression. On the other hand, we have found that induced levels of expression from Pu and P[subscript S] in the presence of YE are partially derepressed in a ptsN mutant of P. putida. PtsN but not Crc seems to directly interfere with XylR activation at target promoters. The effect of the double mutation in ptsN and crc is not the sum of the effects of each independent mutation and suggests that both regulators are elements of a common regulatory pathway. Basal expression levels from these promoters in the absence of inducer are still XylR dependent and are also repressed in the presence of yeast extract. Neither crc nor ptsN could relieve this repression.
ISSN:	0099-2240 1098-5336