Siderophore-mediated signaling regulates virulence factor production in Pseudomonas aeruginosa

Numerous bacteria secrete low molecular weight compounds termed siderophores that have a high affinity for iron ions. Siderophores have a well-documented role as iron-scavenging chemicals, chelating iron ions in the environment whereupon the ferrisiderophores reenter the bacterial cells by means of specific cell-surface receptors. The iron is then released for incorporation into bacterial proteins. Here we show that in addition to its role as an iron-scavenger, the siderophore pyoverdine that is secreted by Pseudomonas aeruginosa regulates the production of at least three virulence factors (exotoxin A, an endoprotease, and pyoverdine itself), which are major contributors to the ability of this bacterium to cause disease. Regulation occurs through a transmembrane signaling system that includes an outer membrane receptor for ferripyoverdine, a signal-transducing protein that is predicted to extend from the periplasm into the cytoplasm, and a sigma factor. Expression of genes that form part of the regulon is triggered by pyoverdine so that this siderophore acts as a signaling molecule to control the production of secreted products. Recognition that a siderophore acts as a signaling molecule has important implications for the understanding of interactions between bacterial cells.

[1]  Herman P. Spaink,et al.  Promoters in the nodulation region of the Rhizobium leguminosarum Sym plasmid pRL1JI , 2004, Plant Molecular Biology.

[2]  H. Cunliffe,et al.  Characterization of an Endoprotease (PrpL) Encoded by a PvdS-Regulated Gene in Pseudomonas aeruginosa , 2001, Infection and Immunity.

[3]  N. A. Whitehead,et al.  Quorum-sensing in Gram-negative bacteria. , 2001, FEMS microbiology reviews.

[4]  I. Lamont,et al.  Involvement of a transformylase enzyme in siderophore synthesis in Pseudomonas aeruginosa. , 2001, Microbiology.

[5]  I. Lamont,et al.  Analysis of Promoters Recognized by PvdS, an Extracytoplasmic-Function Sigma Factor Protein fromPseudomonas aeruginosa , 2001, Journal of bacteriology.

[6]  T. J. Brickman,et al.  Transcriptional Activation of BordetellaAlcaligin Siderophore Genes Requires the AlcR Regulator with Alcaligin as Inducer , 2001, Journal of bacteriology.

[7]  F. Pattus,et al.  Iron‐free pyoverdin binds to its outer membrane receptor FpvA in Pseudomonas aeruginosa: a new mechanism for membrane iron transport , 2001, Molecular microbiology.

[8]  S. Lory,et al.  Complete genome sequence of Pseudomonas aeruginosa PAO1, an opportunistic pathogen , 2000, Nature.

[9]  E. Greenberg,et al.  Acyl-homoserine lactone quorum sensing in gram-negative bacteria: a signaling mechanism involved in associations with higher organisms. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[10]  I. Lamont,et al.  Characterization of an ECF sigma factor protein from Pseudomonas aeruginosa. , 2000, Biochemical and biophysical research communications.

[11]  J. Lyczak,et al.  Establishment of Pseudomonas aeruginosa infection: lessons from a versatile opportunist. , 2000, Microbes and infection.

[12]  H. Nitanai,et al.  Impact of Siderophore Production onPseudomonas aeruginosa Infections in Immunosuppressed Mice , 2000, Infection and Immunity.

[13]  V. de Lorenzo,et al.  Functional Analysis of PvdS, an Iron Starvation Sigma Factor of Pseudomonas aeruginosa , 2000, Journal of bacteriology.

[14]  V. Braun,et al.  Surface Signaling in Ferric Citrate Transport Gene Induction: Interaction of the FecA, FecR, and FecI Regulatory Proteins , 2000, Journal of bacteriology.

[15]  C. Ratledge,et al.  Iron metabolism in pathogenic bacteria. , 2000, Annual review of microbiology.

[16]  K. M. Lee,et al.  Identification of genes controlled by quorum sensing in Pseudomonas aeruginosa. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[17]  V. Braun,et al.  Bacterial solutions to the iron-supply problem. , 1999, Trends in biochemical sciences.

[18]  D. Expert WITHHOLDING AND EXCHANGING IRON: Interactions Between Erwinia spp. and Their Plant Hosts. , 1999, Annual review of phytopathology.

[19]  C. Reimmann,et al.  Dihydroaeruginoic acid synthetase and pyochelin synthetase, products of the pchEF genes, are induced by extracellular pyochelin in Pseudomonas aeruginosa. , 1998, Microbiology.

[20]  J. M. Meyer,et al.  Quorum-sensing and siderophore biosynthesis in Pseudomonas aeruginosa: lasR/lasI mutants exhibit reduced pyoverdine biosynthesis. , 1998, FEMS microbiology letters.

[21]  H. Schweizer,et al.  A broad-host-range Flp-FRT recombination system for site-specific excision of chromosomally-located DNA sequences: application for isolation of unmarked Pseudomonas aeruginosa mutants. , 1998, Gene.

[22]  V. Braun,et al.  Ferric Citrate Transport of Escherichia coli: Functional Regions of the FecR Transmembrane Regulatory Protein , 1998, Journal of Bacteriology.

[23]  J. Heesemann,et al.  The Yersiniabactin Biosynthetic Gene Cluster of Yersinia enterocolitica: Organization and Siderophore-Dependent Regulation , 1998, Journal of bacteriology.

[24]  J. Shapiro Thinking about bacterial populations as multicellular organisms. , 1998, Annual review of microbiology.

[25]  J. H. Crosa Signal transduction and transcriptional and posttranscriptional control of iron-regulated genes in bacteria , 1997, Microbiology and molecular biology reviews : MMBR.

[26]  Thomas L. Madden,et al.  Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. , 1997, Nucleic acids research.

[27]  P. Cornelis,et al.  Use of siderophores to type pseudomonads: the three Pseudomonas aeruginosa pyoverdine systems. , 1997, Microbiology.

[28]  Regulation of the iron uptake genes in Pseudomonas fluorescens M114 by pseudobactin M114: the pbrA sigma factor gene does not mediate the siderophore regulatory response. , 1996, FEMS microbiology letters.

[29]  H. Cunliffe,et al.  Exotoxin A production in Pseudomonas aeruginosa requires the iron‐regulated pvdS gene encoding an alternative sigma factor , 1996, Molecular microbiology.

[30]  P. Visca,et al.  Iron-regulated transcription of the pvdA gene in Pseudomonas aeruginosa: effect of Fur and PvdS on promoter activity , 1996, Journal of bacteriology.

[31]  A. Neely,et al.  Pyoverdin is essential for virulence of Pseudomonas aeruginosa , 1996, Infection and immunity.

[32]  P. Weisbeek,et al.  Gene regulation of siderophore‐mediated iron acquisition in Pseudomonas: not only the Fur repressor , 1995, Molecular microbiology.

[33]  H. Cunliffe,et al.  Cloning and characterization of pvdS, a gene required for pyoverdine synthesis in Pseudomonas aeruginosa: PvdS is probably an alternative sigma factor , 1995, Journal of bacteriology.

[34]  J. Thompson,et al.  CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. , 1994, Nucleic acids research.

[35]  M. Guerinot Microbial iron transport. , 1994, Annual review of microbiology.

[36]  M. Gambello,et al.  Expression of Pseudomonas aeruginosa virulence genes requires cell-to-cell communication. , 1993, Science.

[37]  T. Kuo,et al.  A simple and rapid method for the preparation of gram-negative bacterial genomic DNA. , 1993, Nucleic acids research.

[38]  R. Fick,et al.  Pseudomonas aeruginosa, the opportunist : pathogenesis and disease , 1993 .

[39]  J. Marks,et al.  Siderophore presence in sputa of cystic fibrosis patients , 1991 .

[40]  R. Rao,et al.  A thiostrepton-inducible expression vector for use in Streptomyces spp. , 1991, Gene.

[41]  H. Sambrook Molecular cloning : a laboratory manual. Cold Spring Harbor, NY , 1989 .

[42]  A. Pühler,et al.  Plasmid vectors for the genetic analysis and manipulation of rhizobia and other gram-negative bacteria. , 1986, Methods in enzymology.

[43]  J. Sambrook,et al.  Molecular Cloning: A Laboratory Manual , 2001 .

[44]  Jeffrey H. Miller Experiments in molecular genetics , 1972 .

[45]  S. B. Needleman,et al.  A general method applicable to the search for similarities in the amino acid sequence of two proteins. , 1970, Journal of molecular biology.

[46]  King Eo,et al.  Two simple media for the demonstration of pyocyanin and fluorescin. , 1954 .