PMID:19114474

Citation	de Bruijn, I and Raaijmakers, JM (2009) Regulation of cyclic lipopeptide biosynthesis in Pseudomonas fluorescens by the ClpP protease. J. Bacteriol. 191:1910-23
Abstract	Cyclic lipopeptides produced by Pseudomonas species exhibit potent surfactant and broad-spectrum antibiotic properties. Their biosynthesis is governed by large multimodular nonribosomal peptide synthetases, but little is known about the genetic regulatory network. This study provides, for the first time, evidence that the serine protease ClpP regulates the biosynthesis of massetolides, cyclic lipopeptides involved in swarming motility, biofilm formation, and antimicrobial activity of Pseudomonas fluorescens SS101. The results show that ClpP affects the expression of luxR(mA), the transcriptional regulator of the massetolide biosynthesis genes massABC, thereby regulating biofilm formation and swarming motility of P. fluorescens SS101. Transcription of luxR(mA) was significantly repressed in the clpP mutant, and introduction of luxR(mA) restored, in part, massetolide biosynthesis and swarming motility of the clpP mutant. Site-directed mutagenesis and expression analyses indicated that the chaperone subunit ClpX and the Lon protease are not involved in regulation of massetolide biosynthesis and are transcribed independently of clpP. Addition of Casamino Acids enhanced the transcription of luxR(mA) and massABC in the clpP mutant, leading to a partial rescue of massetolide production and swarming motility. The results further suggested that, at the transcriptional level, ClpP-mediated regulation of massetolide biosynthesis operates independently of regulation by the GacA/GacS two-component system. The role of amino acid metabolism and the putative mechanisms underlying ClpP-mediated regulation of cyclic lipopeptide biosynthesis, swarming motility, and growth in P. fluorescens are discussed.
Links	PubMed PMC2648375 Online version:10.1128/JB.01558-08
Keywords	Bacterial Proteins/genetics; Bacterial Proteins/metabolism; Biofilms; Endopeptidase Clp/genetics; Endopeptidase Clp/metabolism; Gene Expression Regulation, Bacterial; Lipopeptides/biosynthesis; Molecular Sequence Data; Pseudomonas fluorescens/enzymology; Pseudomonas fluorescens/genetics; Pseudomonas fluorescens/physiology; Repressor Proteins/genetics; Repressor Proteins/metabolism; Trans-Activators/genetics; Trans-Activators/metabolism; Transcription, Genetic
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Annotations

Gene product	Qualifier	GO Term	Evidence Code	with/from	Aspect	Extension	Notes	Status
PSEFL:I4K5Z9	Contributes to	GO:0042158: lipoprotein biosynthetic process	ECO:0000315:		P		Fig. 1B shows HPLC analysis of massetolide A production of WT and clpP mutants. Strains lacking clpP are unable to produce massetolide A; however, this is massetolide A production is restored upon complementation.	complete CACAO 10835
PSEFL:I4K5Z9	Contributes to	GO:0071978: bacterial-type flagellum-dependent swarming motility	ECO:0000315:		P		Fig. 3D shows the swarming motility of WT and clpP mutants. Strains lacking clpP are unable to swarm; however, this is restored by complementation.	complete CACAO 10836
PSEFL:I4K5Z9		GO:1900192: positive regulation of single-species biofilm formation	ECO:0000315:		P		Fig. 3C shows the biofilm formation of WT and clpP mutants. The ability to form a biofilm in strains lacking clpP is reduced. Biofilm formation was restored upon complementation.	complete CACAO 10837
PSEFL:I4K5Z9	involved_in	GO:1900192: positive regulation of single-species biofilm formation	ECO:0000315: mutant phenotype evidence used in manual assertion		P		Seeded From UniProt	complete

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