An aromatic effector specificity mutant of the transcriptional regulator DmpR overcomes the growth constraints of Pseudomonas sp. strain CF600 on para-substituted methylphenols.
The pVI150 catabolic plasmid of Pseudomonas sp. strain CF600 carries the dmp system, which comprises the divergently transcribed dmpR gene and the dmp operon coding for the catabolic enzymes required for growth on (methyl)phenols. The constitutively expressed DmpR transcriptional activator positively controls the expression of the RpoN-dependent dmp operon promoter in the presence of the aromatic effector in the growth medium. However, the magnitude of the transcriptional response differs depending on the position of the methyl substituent on the aromatic ring. Experiments involving an elevated copy number of the dmp system demonstrate that growth on para-substituted methylphenols is limited by the level of the catabolic enzymes. An effector specificity mutant of DmpR, DmpR-E135K, that responded to the presence of 4-ethylphenol, a noneffector of the wild-type protein, was isolated by genetic selection. The single point mutation in DmpR-E135K, which results in a Glu-to-Lys change in residue 135, also results in a regulator with enhanced recognition of para-substituted methylphenols. The DmpR-E135K mutation, when introduced into the wild-type strain, confers enhanced utilization of the para-substituted methylphenols. These experiments demonstrate that the aromatic effector activation of wild-type DmpR by the para-substituted methylphenols is a major factor limiting the catabolism of these compounds.
Pavel
H
H
Department of Cell and Molecular Biology, Umeå University, Sweden.
Forsman
M
M
Shingler
V
V
D001426:Bacterial Proteins / Q000235:genetics*; D001673:Biodegradation, Environmental; D002245:Carbon Dioxide / Q000378:metabolism; D003408:Cresols / Q000378:metabolism; D018628:Gene Dosage; D015964:Gene Expression Regulation, Bacterial / Q000187:drug effects; D017930:Genes, Reporter; D006899:Mixed Function Oxygenases / Q000096:biosynthesis; D009154:Mutation; D019800:Phenol; D010636:Phenols / Q000378:metabolism* / Q000494:pharmacology*; D011549:Pseudomonas / Q000145:classification / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism; D011995:Recombination, Genetic; D015183:Restriction Mapping; D012641:Selection, Genetic; D015534:Trans-Activators / Q000235:genetics*; D014158:Transcription, Genetic / Q000187:drug effects; D014992:Xylenes / Q000378:metabolism
D001426:Bacterial Proteins; D003408:Cresols; C080133:DmpR protein, Pseudomonas; D010636:Phenols; D015534:Trans-Activators; D014992:Xylenes; D002245:Carbon Dioxide; C032538:4-cresol; D019800:Phenol; C063628:3,4-dimethylphenol; C042291:4-ethylphenol; D006899:Mixed Function Oxygenases; C022336:phenol 2-monooxygenase