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A Serratia marcescens PigP homolog controls prodigiosin biosynthesis, swarming motility and hemolysis and is regulated by cAMP-CRP and HexS.

Shanks RM, Lahr RM, Stella NA, Arena KE, Brothers KM, Kwak DH, Liu X, Kalivoda EJ - PLoS ONE (2013)

Bottom Line: Transcriptional analysis and electrophoretic mobility shift assays place PigP in a regulatory pathway with upstream regulators CRP and HexS.The data from this study identifies a positive regulator of serratamolide production, describes novel roles for the PigP transcription factor, shows for the first time that PigP directly regulates the pigment biosynthetic operon, and identifies upstream regulators of pigP.This study suggests that PigP is important for the ability of S. marcescens to compete in the environment.

View Article: PubMed Central - PubMed

Affiliation: Charles T Campbell Laboratory of Ophthalmic Microbiology, Department of Ophthalmology, University of Pittsburgh Eye Center, Pittsburgh, Pennsylvania, United States of America.

ABSTRACT
Swarming motility and hemolysis are virulence-associated determinants for a wide array of pathogenic bacteria. The broad host-range opportunistic pathogen Serratia marcescens produces serratamolide, a small cyclic amino-lipid, that promotes swarming motility and hemolysis. Serratamolide is negatively regulated by the transcription factors HexS and CRP. Positive regulators of serratamolide production are unknown. Similar to serratamolide, the antibiotic pigment, prodigiosin, is regulated by temperature, growth phase, HexS, and CRP. Because of this co-regulation, we tested the hypothesis that a homolog of the PigP transcription factor of the atypical Serratia species ATCC 39006, which positively regulates prodigiosin biosynthesis, is also a positive regulator of serratamolide production in S. marcescens. Mutation of pigP in clinical, environmental, and laboratory strains of S. marcescens conferred pleiotropic phenotypes including the loss of swarming motility, hemolysis, and severely reduced prodigiosin and serratamolide synthesis. Transcriptional analysis and electrophoretic mobility shift assays place PigP in a regulatory pathway with upstream regulators CRP and HexS. The data from this study identifies a positive regulator of serratamolide production, describes novel roles for the PigP transcription factor, shows for the first time that PigP directly regulates the pigment biosynthetic operon, and identifies upstream regulators of pigP. This study suggests that PigP is important for the ability of S. marcescens to compete in the environment.

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PigP is necessary for swarming motility and serratamolide production, but not swimming motility.A. Swarming motility plates show that the pigP mutant is defective in surface motility. Mutation of the serratamolide inhibitor hexS restores swarming to the pigP mutant (hexS pigP), and hexS requires serratamolide to swarm (hexS swrW). B. The pigP swarming defect can be complemented by the wild-type pigP gene provided in trans. Vector refers to pMQ132, and ppigP is pMQ221. C. Swimming motility plates show similar zones (in all cases “zones” indicates the measurement from the edge of the colony to the outer edge of the observed phenotype in mm) between the WT and pigP mutant. D. Surfactant zones are absent in strains without pigP and the serratamolide biosynthetic gene swrW. Mutation of hexS restores surfactant zones to the pigP mutant. N ≥6 biological replicates per strain. E. Arabinose-induced expression of swrW is sufficient to restore swarming to a swrW and pigP mutant. pMQ200+ swrW refers to pMQ368. F. Purified serratamolide is sufficient to restore swarming to different strain backgrounds with pigP mutations, whereas the serratamolide vehicle, DMSO, is not.
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pone-0057634-g005: PigP is necessary for swarming motility and serratamolide production, but not swimming motility.A. Swarming motility plates show that the pigP mutant is defective in surface motility. Mutation of the serratamolide inhibitor hexS restores swarming to the pigP mutant (hexS pigP), and hexS requires serratamolide to swarm (hexS swrW). B. The pigP swarming defect can be complemented by the wild-type pigP gene provided in trans. Vector refers to pMQ132, and ppigP is pMQ221. C. Swimming motility plates show similar zones (in all cases “zones” indicates the measurement from the edge of the colony to the outer edge of the observed phenotype in mm) between the WT and pigP mutant. D. Surfactant zones are absent in strains without pigP and the serratamolide biosynthetic gene swrW. Mutation of hexS restores surfactant zones to the pigP mutant. N ≥6 biological replicates per strain. E. Arabinose-induced expression of swrW is sufficient to restore swarming to a swrW and pigP mutant. pMQ200+ swrW refers to pMQ368. F. Purified serratamolide is sufficient to restore swarming to different strain backgrounds with pigP mutations, whereas the serratamolide vehicle, DMSO, is not.

Mentions: Swarming motility over the surface is a virulence associated group behavior [65]–[67]. We observed that the ΔpigP strain (CMS1713) was unable to swarm (0%, n = 15, performed on 3 different days), whereas WT was capable of swarming (100%, n = 15) (Figure 5A). The pigP mutant swarming deficiency could be complemented by WT pigP on a plasmid (Figure 5B).


A Serratia marcescens PigP homolog controls prodigiosin biosynthesis, swarming motility and hemolysis and is regulated by cAMP-CRP and HexS.

Shanks RM, Lahr RM, Stella NA, Arena KE, Brothers KM, Kwak DH, Liu X, Kalivoda EJ - PLoS ONE (2013)

PigP is necessary for swarming motility and serratamolide production, but not swimming motility.A. Swarming motility plates show that the pigP mutant is defective in surface motility. Mutation of the serratamolide inhibitor hexS restores swarming to the pigP mutant (hexS pigP), and hexS requires serratamolide to swarm (hexS swrW). B. The pigP swarming defect can be complemented by the wild-type pigP gene provided in trans. Vector refers to pMQ132, and ppigP is pMQ221. C. Swimming motility plates show similar zones (in all cases “zones” indicates the measurement from the edge of the colony to the outer edge of the observed phenotype in mm) between the WT and pigP mutant. D. Surfactant zones are absent in strains without pigP and the serratamolide biosynthetic gene swrW. Mutation of hexS restores surfactant zones to the pigP mutant. N ≥6 biological replicates per strain. E. Arabinose-induced expression of swrW is sufficient to restore swarming to a swrW and pigP mutant. pMQ200+ swrW refers to pMQ368. F. Purified serratamolide is sufficient to restore swarming to different strain backgrounds with pigP mutations, whereas the serratamolide vehicle, DMSO, is not.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3585978&req=5

pone-0057634-g005: PigP is necessary for swarming motility and serratamolide production, but not swimming motility.A. Swarming motility plates show that the pigP mutant is defective in surface motility. Mutation of the serratamolide inhibitor hexS restores swarming to the pigP mutant (hexS pigP), and hexS requires serratamolide to swarm (hexS swrW). B. The pigP swarming defect can be complemented by the wild-type pigP gene provided in trans. Vector refers to pMQ132, and ppigP is pMQ221. C. Swimming motility plates show similar zones (in all cases “zones” indicates the measurement from the edge of the colony to the outer edge of the observed phenotype in mm) between the WT and pigP mutant. D. Surfactant zones are absent in strains without pigP and the serratamolide biosynthetic gene swrW. Mutation of hexS restores surfactant zones to the pigP mutant. N ≥6 biological replicates per strain. E. Arabinose-induced expression of swrW is sufficient to restore swarming to a swrW and pigP mutant. pMQ200+ swrW refers to pMQ368. F. Purified serratamolide is sufficient to restore swarming to different strain backgrounds with pigP mutations, whereas the serratamolide vehicle, DMSO, is not.
Mentions: Swarming motility over the surface is a virulence associated group behavior [65]–[67]. We observed that the ΔpigP strain (CMS1713) was unable to swarm (0%, n = 15, performed on 3 different days), whereas WT was capable of swarming (100%, n = 15) (Figure 5A). The pigP mutant swarming deficiency could be complemented by WT pigP on a plasmid (Figure 5B).

Bottom Line: Transcriptional analysis and electrophoretic mobility shift assays place PigP in a regulatory pathway with upstream regulators CRP and HexS.The data from this study identifies a positive regulator of serratamolide production, describes novel roles for the PigP transcription factor, shows for the first time that PigP directly regulates the pigment biosynthetic operon, and identifies upstream regulators of pigP.This study suggests that PigP is important for the ability of S. marcescens to compete in the environment.

View Article: PubMed Central - PubMed

Affiliation: Charles T Campbell Laboratory of Ophthalmic Microbiology, Department of Ophthalmology, University of Pittsburgh Eye Center, Pittsburgh, Pennsylvania, United States of America.

ABSTRACT
Swarming motility and hemolysis are virulence-associated determinants for a wide array of pathogenic bacteria. The broad host-range opportunistic pathogen Serratia marcescens produces serratamolide, a small cyclic amino-lipid, that promotes swarming motility and hemolysis. Serratamolide is negatively regulated by the transcription factors HexS and CRP. Positive regulators of serratamolide production are unknown. Similar to serratamolide, the antibiotic pigment, prodigiosin, is regulated by temperature, growth phase, HexS, and CRP. Because of this co-regulation, we tested the hypothesis that a homolog of the PigP transcription factor of the atypical Serratia species ATCC 39006, which positively regulates prodigiosin biosynthesis, is also a positive regulator of serratamolide production in S. marcescens. Mutation of pigP in clinical, environmental, and laboratory strains of S. marcescens conferred pleiotropic phenotypes including the loss of swarming motility, hemolysis, and severely reduced prodigiosin and serratamolide synthesis. Transcriptional analysis and electrophoretic mobility shift assays place PigP in a regulatory pathway with upstream regulators CRP and HexS. The data from this study identifies a positive regulator of serratamolide production, describes novel roles for the PigP transcription factor, shows for the first time that PigP directly regulates the pigment biosynthetic operon, and identifies upstream regulators of pigP. This study suggests that PigP is important for the ability of S. marcescens to compete in the environment.

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Related in: MedlinePlus