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Journal of Bacteriology, December 2006, p. 8283-8293, Vol. 188, No. 23
0021-9193/06/$08.00+0     doi:10.1128/JB.00893-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Pip, a Novel Activator of Phenazine Biosynthesis in Pseudomonas chlororaphis PCL1391{triangledown} ,{dagger}

Geneviève Girard,1 Sharief Barends,2 Sébastien Rigali,2 E. Tjeerd van Rij,1 Ben J. J. Lugtenberg,1 and Guido V. Bloemberg1*

Leiden University, Institute of Biology, Clusius Laboratory, Wassenaarseweg 64, 2333 AL Leiden, The Netherlands,1 Leiden University, Leiden Institute of Chemistry, Department of Molecular Genetics, P.O. Box 9502, 2300 RA Leiden, The Netherlands2

Received 21 June 2006/ Accepted 14 September 2006

Secondary metabolites are important factors for interactions between bacteria and other organisms. Pseudomonas chlororaphis PCL1391 produces the antifungal secondary metabolite phenazine-1-carboxamide (PCN) that inhibits growth of Fusarium oxysporum f. sp. radius lycopersici the causative agent of tomato foot and root rot. Our previous work unraveled a cascade of genes regulating the PCN biosynthesis operon, phzABCDEFGH. Via a genetic screen, we identify in this study a novel TetR/AcrR regulator, named Pip (phenazine inducing protein), which is essential for PCN biosynthesis. A combination of a phenotypical characterization of a pip mutant, in trans complementation assays of various mutant strains, and electrophoretic mobility shift assays identified Pip as the fifth DNA-binding protein so far involved in regulation of PCN biosynthesis. In this regulatory pathway, Pip is positioned downstream of PsrA (Pseudomonas sigma factor regulator) and the stationary-phase sigma factor RpoS, while it is upstream of the quorum-sensing system PhzI/PhzR. These findings provide further evidence that the path leading to the expression of secondary metabolism gene clusters in Pseudomonas species is highly complex.


* Corresponding author. Mailing address: Leiden University, Institute of Biology, Clusius Laboratory, Wassenaarseweg 64, 2333 AL Leiden, The Netherlands. Phone: 31 71 527 5056. Fax: 31 71 527 4999. E-mail: g.v.bloemberg{at}biology.leidenuniv.nl.

{triangledown} Published ahead of print on 22 September 2006.

{dagger} Supplemental material for this article may be found at http://jb.asm.org/.


Journal of Bacteriology, December 2006, p. 8283-8293, Vol. 188, No. 23
0021-9193/06/$08.00+0     doi:10.1128/JB.00893-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.




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