Regulatory roles of psrA and rpoS in phenazine-1-carboxamide synthesis by Pseudomonas chlororaphis PCL1391.
Production of the secondary metabolite phenazine-1-carboxamide (PCN) by Pseudomonas chlororaphis PCL1391 is crucial for biocontrol activity against the phytopathogen Fusarium oxysporum f. sp. radicis lycopersici on tomato. Regulation of PCN production involves the two-component signalling system GacS/GacA, the quorum-sensing system PhzI/PhzR and the regulator PsrA. This paper reports that a functional rpoS is required for optimal PCN and N-hexanoyl-L-homoserine lactone (C(6)-HSL) production. Constitutive expression of rpoS is able to complement partially the defect of a psrA mutant for PCN and N-acylhomoserine lactone production. Western blotting shows that rpoS is regulated by gacS. Altogether, these results suggest the existence of a cascade consisting of gacS/gacA upstream of psrA and rpoS, which influence expression of phzI/phzR. Overproduction of phzR complements the effects on PCN and C(6)-HSL production of all mutations tested in the regulatory cascade, which shows that a functional quorum-sensing system is essential and sufficient for PCN synthesis. In addition, the relative amounts of PCN, phenazine-1-carboxylic acid and C(6)-HSL produced by rpoS and psrA mutants harbouring a constitutively expressed phzR indicate an even more complex network of interactions, probably involving other genes. Preliminary microarray analyses of the transcriptomics of the rpoS and psrA mutants support the model of regulation described in this study and allow identification of new genes that might be involved in secondary metabolism.
Girard
Geneviève
G
Leiden University, Institute of Biology (IBL), Clusius Laboratory, Wassenaarseweg 64, 2333AL Leiden, the Netherlands.
van Rij
E Tjeerd
ET
Leiden University, Institute of Biology (IBL), Clusius Laboratory, Wassenaarseweg 64, 2333AL Leiden, the Netherlands.
Lugtenberg
Ben J J
BJJ
Leiden University, Institute of Biology (IBL), Clusius Laboratory, Wassenaarseweg 64, 2333AL Leiden, the Netherlands.
Bloemberg
Guido V
GV
Leiden University, Institute of Biology (IBL), Clusius Laboratory, Wassenaarseweg 64, 2333AL Leiden, the Netherlands.
D015107:4-Butyrolactone / Q000031:analogs & derivatives / Q000378:metabolism; D000935:Antifungal Agents / Q000378:metabolism; D001426:Bacterial Proteins / Q000235:genetics / Q000502:physiology*; D004268:DNA-Binding Proteins / Q000235:genetics / Q000502:physiology*; D005786:Gene Expression Regulation; D005798:Genes, Bacterial; D009876:Operon; D010619:Phenazines / Q000378:metabolism*; D011549:Pseudomonas / Q000235:genetics / Q000378:metabolism*; D018085:Regulon; D012808:Sigma Factor / Q000235:genetics / Q000502:physiology*; D015534:Trans-Activators / Q000235:genetics / Q000502:physiology; D014157:Transcription Factors / Q000235:genetics / Q000502:physiology*
D000935:Antifungal Agents; D001426:Bacterial Proteins; D004268:DNA-Binding Proteins; C091100:GacA protein, Bacteria; C104908:N-hexanoyl-L-homoserine lactone; D010619:Phenazines; C088011:PhzR protein, Pseudomonas; C430183:PsrA protein, Pseudomonas; D012808:Sigma Factor; D015534:Trans-Activators; D014157:Transcription Factors; C420923:phenazine-1-carboxamide; C069567:sigma factor KatF protein, Bacteria; C074165:lemA protein, bacterial; D015107:4-Butyrolactone