Transcriptional control of quorum sensing and associated metabolic interactions in Pseudomonas syringae strain B728a

Pseudomonas syringae pv. syringae cell densities fluctuate regularly during host plant colonization. Previously we identified nine genes dependent on the quorum‐sensing‐associated luxR homolog ahlR during epiphytic and apoplastic stages of host colonization. Yet their contributions to host colonization remain obscure, despite ahlR regulon presence within and beyond the P. syringae pan‐genome. To elucidate AhIR regulon member functions, we characterized their regulation, interactions with each other, and contributions to the metabolome. We report Psyr_1625, encoding a functional pyruvate deydrogenase‐E1 subunit PdhQ, is required to prevent the accumulation of pyruvate in rich media. Furthermore it is exquisitely regulated by both repression of its own promoter by QrpR within a novel clade of the MarR regulator family, and co‐transcription on a 5kb transcript originating from the AhlR‐driven ahlI promoter, that reads over ahlR and qrpR. Metabolites accumulated during expression of the second AhlR‐driven operon (Psyr_1620‐1616, paoABCDE), only in a pdhQ mutant background, in addition to pyruvate, are herein associated with derepression of QrpR‐repressed pdhQ. AHL signaling, QrpR, and transcriptional read‐through events integrate to ensure AHL‐dependent expression of a novel metabolism in anticipation of environmental stress, while minimizing endogenously generated cytotoxicity.

[1]  Tomoko Suzuki,et al.  Characterization of quorum sensing-controlled transcriptional regulator MarR and Rieske (2Fe-2S) cluster-containing protein (Orf5), which are involved in resistance to environmental stresses in Pseudomonas syringae pv. tabaci 6605. , 2015, Molecular plant pathology.

[2]  S. Lund,et al.  Transcriptional Analysis of the Global Regulatory Networks Active in Pseudomonas syringae during Leaf Colonization , 2014, mBio.

[3]  Yuval Hart,et al.  The utility of paradoxical components in biological circuits. , 2013, Molecular cell.

[4]  Steven P. Lund,et al.  Transcriptional responses of Pseudomonas syringae to growth in epiphytic versus apoplastic leaf sites , 2013, Proceedings of the National Academy of Sciences.

[5]  E. Greenberg,et al.  Strain-dependent diversity in the Pseudomonas aeruginosa quorum-sensing regulon , 2012, Proceedings of the National Academy of Sciences.

[6]  C. Kuttler,et al.  Stochastic modeling of Pseudomonas syringae growth in the phyllosphere. , 2012, Mathematical biosciences.

[7]  E. Greenberg,et al.  Antisense RNA that affects Rhodopseudomonas palustris quorum-sensing signal receptor expression , 2012, Proceedings of the National Academy of Sciences.

[8]  S. Lindow,et al.  XagR, a LuxR homolog, contributes to the virulence of Xanthomonas axonopodis pv. glycines to soybean. , 2012, Molecular plant-microbe interactions : MPMI.

[9]  Peter Williams,et al.  IMG: the integrated microbial genomes database and comparative analysis system , 2011, Nucleic Acids Res..

[10]  Nathan Linial,et al.  ProtoNet 6.0: organizing 10 million protein sequences in a compact hierarchical family tree , 2011, Nucleic Acids Res..

[11]  Robert D. Finn,et al.  InterPro in 2011: new developments in the family and domain prediction database , 2011, Nucleic acids research.

[12]  Susumu Goto,et al.  KEGG for integration and interpretation of large-scale molecular data sets , 2011, Nucleic Acids Res..

[13]  V. Venturi,et al.  Functional Characterization of the Quorum Sensing Regulator RsaL in the Plant-Beneficial Strain Pseudomonas putida WCS358 , 2011, Applied and Environmental Microbiology.

[14]  Andrew G. Palmer,et al.  Attenuation of virulence in pathogenic bacteria using synthetic quorum-sensing modulators under native conditions on plant hosts. , 2011, ACS chemical biology.

[15]  M. Shahid Mukhtar,et al.  Dynamic Evolution of Pathogenicity Revealed by Sequencing and Comparative Genomics of 19 Pseudomonas syringae Isolates , 2011, PLoS pathogens.

[16]  H. Vogel,et al.  Differences in metabolism between the biofilm and planktonic response to metal stress. , 2011, Journal of proteome research.

[17]  Ian Sillitoe,et al.  Extending CATH: increasing coverage of the protein structure universe and linking structure with function , 2010, Nucleic Acids Res..

[18]  Damian Szklarczyk,et al.  The STRING database in 2011: functional interaction networks of proteins, globally integrated and scored , 2010, Nucleic Acids Res..

[19]  Raymond Lo,et al.  Pseudomonas Genome Database: improved comparative analysis and population genomics capability for Pseudomonas genomes , 2010, Nucleic Acids Res..

[20]  Vittorio Venturi,et al.  The plant pathogen Pseudomonas fuscovaginae contains two conserved quorum sensing systems involved in virulence and negatively regulated by RsaL and the novel regulator RsaM. , 2011, Environmental microbiology.

[21]  S. C. Winans,et al.  LuxR‐type quorum‐sensing regulators that are detached from common scents , 2010, Molecular microbiology.

[22]  S. Lindow,et al.  Interference of quorum sensing in Pseudomonas syringae by bacterial epiphytes that limit iron availability. , 2010, Environmental microbiology.

[23]  Royston Goodacre,et al.  Evaluation of extraction processes for intracellular metabolite profiling of mammalian cells: matching extraction approaches to cell type and metabolite targets , 2010, Metabolomics.

[24]  Bronwyn G. Butcher,et al.  Transcriptome Analysis of Pseudomonas syringae Identifies New Genes, Noncoding RNAs, and Antisense Activity , 2010, Journal of bacteriology.

[25]  Vittorio Venturi,et al.  LuxR-family 'solos': bachelor sensors/regulators of signalling molecules. , 2009, Microbiology.

[26]  Alexander P. Hynes,et al.  Metabolomic Investigation of the Bacterial Response to a Metal Challenge , 2008, Applied and Environmental Microbiology.

[27]  John D. Westbrook,et al.  The Protein Model Portal , 2008, Journal of Structural and Functional Genomics.

[28]  S. Lindow,et al.  Two Dissimilar N-Acyl-Homoserine Lactone Acylases of Pseudomonas syringae Influence Colony and Biofilm Morphology , 2008, Applied and Environmental Microbiology.

[29]  Yu Tanouchi,et al.  Noise Reduction by Diffusional Dissipation in a Minimal Quorum Sensing Motif , 2008, PLoS Comput. Biol..

[30]  R. Goodacre,et al.  Global Metabolic Profiling of Escherichia Coli Cultures: an Evaluation of Methods for Quenching and Extraction of Intracellular Metabolites , 2022 .

[31]  S. Lindow,et al.  Quorum size of Pseudomonas syringae is small and dictated by water availability on the leaf surface , 2008, Proceedings of the National Academy of Sciences.

[32]  B. Bassler,et al.  A negative feedback loop involving small RNAs accelerates Vibrio cholerae's transition out of quorum-sensing mode. , 2008, Genes & development.

[33]  Frank Jordan,et al.  A Dynamic Loop at the Active Center of the Escherichia coli Pyruvate Dehydrogenase Complex E1 Component Modulates Substrate Utilization and Chemical Communication with the E2 Component* , 2007, Journal of Biological Chemistry.

[34]  G. Dulla Bacterial babel: Breaking down quorum sensing cross-talk in the phyllosphere; analysis of the contributions of abiotic and biotic factors on AHL-mediated quorum sensing to epiphytic growth and virulence in Pseudomonas syringae , 2007 .

[35]  C. Fuqua,et al.  The QscR Quorum-Sensing Regulon of Pseudomonas aeruginosa: an Orphan Claims Its Identity , 2006, Journal of bacteriology.

[36]  M. Surette,et al.  Communication in bacteria: an ecological and evolutionary perspective , 2006, Nature Reviews Microbiology.

[37]  H. Mori,et al.  Construction of Escherichia coli K-12 in-frame, single-gene knockout mutants: the Keio collection , 2006, Molecular systems biology.

[38]  S. Sauret-Güeto,et al.  A mutant pyruvate dehydrogenase E1 subunit allows survival of Escherichia coli strains defective in 1‐deoxy‐d‐xylulose 5‐phosphate synthase , 2006, FEBS letters.

[39]  Marc A. Martí-Renom,et al.  MODBASE: a database of annotated comparative protein structure models and associated resources , 2005, Nucleic Acids Res..

[40]  J. Nielsen,et al.  Global metabolite analysis of yeast: evaluation of sample preparation methods , 2005, Yeast.

[41]  S. Lindow,et al.  Quorum sensing regulates exopolysaccharide production, motility, and virulence in Pseudomonas syringae. , 2005, Molecular plant-microbe interactions : MPMI.

[42]  K. Tittmann,et al.  Glutamate 636 of the Escherichia coli Pyruvate Dehydrogenase-E1 Participates in Active Center Communication and Behaves as an Engineered Acetolactate Synthase with Unusual Stereoselectivity* , 2005, Journal of Biological Chemistry.

[43]  Matt Nolan,et al.  Comparison of the complete genome sequences of Pseudomonas syringae pv. syringae B728a and pv. tomato DC3000. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[44]  S. Lindow,et al.  Regulation of AHL production and its contribution to epiphytic fitness in Pseudomonas syringae. , 2004, Molecular plant-microbe interactions : MPMI.

[45]  S. Lindow,et al.  Frequency, Size, and Localization of Bacterial Aggregates on Bean Leaf Surfaces , 2004, Applied and Environmental Microbiology.

[46]  S. Lindow,et al.  Differential survival of solitary and aggregated bacterial cells promotes aggregate formation on leaf surfaces , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[47]  S. Lindow,et al.  Microbiology of the Phyllosphere , 2003, Applied and Environmental Microbiology.

[48]  T. Ferenci,et al.  Global metabolite analysis: the influence of extraction methodology on metabolome profiles of Escherichia coli. , 2003, Analytical biochemistry.

[49]  J. Davison Genetic tools for pseudomonads, rhizobia, and other gram-negative bacteria. , 2002, BioTechniques.

[50]  Thomas D. Schmittgen,et al.  Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. , 2001, Methods.

[51]  J. Leveau,et al.  Improved gfp and inaZ broad-host-range promoter-probe vectors. , 2000, Molecular plant-microbe interactions : MPMI.

[52]  S. Molin,et al.  Detection of N-acylhomoserine lactones in lung tissues of mice infected with Pseudomonas aeruginosa. , 2000, Microbiology.

[53]  B. Wanner,et al.  One-step inactivation of chromosomal genes in Escherichia coli K-12 using PCR products. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[54]  David K. Willis,et al.  Swarming by Pseudomonas syringae B728a Requires gacS (lemA) and gacA but Not the Acyl-Homoserine Lactone Biosynthetic GeneahlI , 1999, Journal of bacteriology.

[55]  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.

[56]  H. Inoue,et al.  Molecular characterization of the mde operon involved in L-methionine catabolism of Pseudomonas putida , 1997, Journal of bacteriology.

[57]  C. Chitnis,et al.  Pseudomonas aeruginosa AlgG is a polymer level alginate C5-mannuronan epimerase , 1994, Journal of bacteriology.

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

[59]  H. Krisch,et al.  In vitro insertional mutagenesis with a selectable DNA fragment. , 1984, Gene.

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

[61]  E. King,et al.  Two simple media for the demonstration of pyocyanin and fluorescin. , 1954, The Journal of laboratory and clinical medicine.