Transcriptional regulation of the pdt gene cluster of Pseudomonas stutzeri KC involves an AraC/XylS family transcriptional activator (PdtC) and the cognate siderophore pyridine-2,6-bis(thiocarboxylic acid).
In order to gain an understanding of the molecular mechanisms dictating production of the siderophore and dechlorination agent pyridine-2,6-bis(thiocarboxylic acid) (PDTC), we have begun characterization of a gene found in the pdt gene cluster of Pseudomonas stutzeri KC predicted to have a regulatory role. That gene product is an AraC family transcriptional activator, PdtC. Quantitative reverse transcription-PCR and expression of transcriptional reporter fusions were used to assess a role for pdtC in the transcription of pdt genes. PdtC and an upstream, promoter-proximal DNA segment were required for wild-type levels of expression from the promoter of a predicted biosynthesis operon (P(pdtF)). At least two other transcriptional units within the pdt cluster were also dependent upon pdtC for expression at wild-type levels. The use of a heterologous, Pseudomonas putida host demonstrated that pdtC and an exogenously added siderophore were necessary and sufficient for expression from the pdtF promoter, i.e., none of the PDTC utilization genes within the pdt cluster were required for transcriptional signaling. Tests using the promoter of the pdtC gene in transcriptional reporter fusions indicated siderophore-dependent negative autoregulation similar to that seen with other AraC-type regulators of siderophore biosynthesis and utilization genes. The data increase the repertoire of siderophore systems known to be regulated by this type of transcriptional activator and have implications for PDTC signaling.
Morales
Sergio E
SE
Department of Microbiology and Molecular Genetics, University of Vermont, 95 Carrigan Dr., Burlington, VT 05405, USA.
Lewis
Thomas A
TA
D051777:AraC Transcription Factor / Q000378:metabolism*; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism; D003470:Culture Media; D015964:Gene Expression Regulation, Bacterial; D005810:Multigene Family; D044223:Pseudomonas stutzeri / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D011725:Pyridines / Q000378:metabolism*; D017262:Siderophores / Q000378:metabolism; D014158:Transcription, Genetic
D051777:AraC Transcription Factor; D001426:Bacterial Proteins; D003470:Culture Media; D011725:Pyridines; D017262:Siderophores; C120960:pyridine-2,6-bis(thiocarboxylic acid)