CodY of Streptococcus pneumoniae: Link between Nutritional Gene Regulation and Colonization

ABSTRACT CodY is a nutritional regulator mainly involved in amino acid metabolism. It has been extensively studied in Bacillus subtilis and Lactococcus lactis. We investigated the role of CodY in gene regulation and virulence of the human pathogen Streptococcus pneumoniae. We constructed a codY mutant and examined the effect on gene and protein expression by microarray and two-dimensional differential gel electrophoresis analysis. The pneumococcal CodY regulon was found to consist predominantly of genes involved in amino acid metabolism but also several other cellular processes, such as carbon metabolism and iron uptake. By means of electrophoretic mobility shift assays and DNA footprinting, we showed that most of the targets identified are under the direct control of CodY. By mutating DNA predicted to represent the CodY box based on the L. lactis consensus, we demonstrated that this sequence is indeed required for in vitro DNA binding to target promoters. Similar to L. lactis, DNA binding of CodY was enhanced in the presence of branched-chain amino acids, but not by GTP. We observed in experimental mouse models that codY is transcribed in the murine nasopharynx and lungs and is specifically required for colonization. This finding was underscored by the diminished ability of the codY mutant to adhere to nasopharyngeal cells in vitro. Furthermore, we found that pcpA, activated by CodY, is required for adherence to nasopharyngeal cells, suggesting a direct link between nutritional regulation and adherence. In conclusion, pneumococcal CodY predominantly regulates genes involved in amino acid metabolism and contributes to the early stages of infection, i.e., colonization of the nasopharynx.

[1]  M. Egmont-Petersen,et al.  Analysis of the In Vitro Transcriptional Response of Human Pharyngeal Epithelial Cells to Adherent Streptococcus pneumoniae: Evidence for a Distinct Response to Encapsulated Strains , 2007, Infection and Immunity.

[2]  P. Andrew,et al.  Characterization of relA and codY mutants of Listeria monocytogenes: identification of the CodY regulon and its role in virulence , 2007, Molecular microbiology.

[3]  Oscar P. Kuipers,et al.  Development of Genomic Array Footprinting for Identification of Conditionally Essential Genes in Streptococcus pneumoniae , 2007, Applied and Environmental Microbiology.

[4]  O. Kuipers,et al.  Regulation of Gene Expression in Streptococcus pneumoniae by Response Regulator 09 Is Strain Dependent , 2006, Journal of bacteriology.

[5]  A. Sonenshein,et al.  Positive regulation of Bacillus subtilis ackA by CodY and CcpA: establishing a potential hierarchy in carbon flow , 2006, Molecular microbiology.

[6]  J. Glass,et al.  Genome Sequence of Avery's Virulent Serotype 2 Strain D39 of Streptococcus pneumoniae and Comparison with That of Unencapsulated Laboratory Strain R6 , 2006, Journal of bacteriology.

[7]  O. Kuipers,et al.  Regulation of Glutamine and Glutamate Metabolism by GlnR and GlnA in Streptococcus pneumoniae* , 2006, Journal of Biological Chemistry.

[8]  H. Malke,et al.  Linking the nutritional status of Streptococcus pyogenes to alteration of transcriptional gene expression: the action of CodY and RelA. , 2006, International journal of medical microbiology : IJMM.

[9]  Oscar P. Kuipers,et al.  BAGEL: a web-based bacteriocin genome mining tool , 2006, Nucleic Acids Res..

[10]  P. Joseph,et al.  The Structure of CodY, a GTP- and Isoleucine-responsive Regulator of Stationary Phase and Virulence in Gram-positive Bacteria* , 2006, Journal of Biological Chemistry.

[11]  D. Briles,et al.  Mn2+-Dependent Regulation of Multiple Genes in Streptococcus pneumoniae through PsaR and the Resultant Impact on Virulence , 2006, Infection and Immunity.

[12]  Dipankar Chatterji,et al.  ppGpp: stringent response and survival. , 2006, Journal of microbiology.

[13]  M. Page,et al.  Global Transcriptome Analysis of the Responses of a Fluoroquinolone-Resistant Streptococcus pneumoniae Mutant and Its Parent to Ciprofloxacin , 2006, Antimicrobial Agents and Chemotherapy.

[14]  N. Pons,et al.  Overall control of nitrogen metabolism in Lactococcus lactis by CodY, and possible models for CodY regulation in Firmicutes. , 2005, Microbiology.

[15]  P. Joseph,et al.  A Region of Bacillus subtilis CodY Protein Required for Interaction with DNA , 2005, Journal of bacteriology.

[16]  Yasutaro Fujita,et al.  Elaborate transcription regulation of the Bacillus subtilis ilv‐leu operon involved in the biosynthesis of branched‐chain amino acids through global regulators of CcpA, CodY and TnrA , 2005, Molecular microbiology.

[17]  Harma A. Karsens,et al.  A generally applicable validation scheme for the assessment of factors involved in reproducibility and quality of DNA-microarray data , 2005, BMC Genomics.

[18]  A. Sonenshein CodY, a global regulator of stationary phase and virulence in Gram-positive bacteria. , 2005, Current opinion in microbiology.

[19]  O. Kuipers,et al.  Probing Direct Interactions between CodY and the oppD Promoter of Lactococcus lactis , 2005, Journal of bacteriology.

[20]  R. de Groot,et al.  Development of antibodies against pneumococcal proteins alpha-enolase, immunoglobulin A1 protease, streptococcal lipoprotein rotamase A, and putative proteinase maturation protein A in relation to pneumococcal carriage and Otitis Media. , 2004, Vaccine.

[21]  A. Sonenshein,et al.  Activation of the Bacillus subtilis global regulator CodY by direct interaction with branched‐chain amino acids , 2004, Molecular microbiology.

[22]  P. Renault,et al.  Intracellular effectors regulating the activity of the Lactococcus lactis CodY pleiotropic transcription regulator , 2004, Molecular microbiology.

[23]  J. Claverys,et al.  The Ami-AliA/AliB Permease of Streptococcus pneumoniae Is Involved in Nasopharyngeal Colonization but Not in Invasive Disease , 2004, Infection and Immunity.

[24]  R. de Groot,et al.  Streptococcus pneumoniae colonisation: the key to pneumococcal disease. , 2004, The Lancet. Infectious diseases.

[25]  Sacha A. F. T. van Hijum,et al.  PreP: gene expression data pre-processing , 2003, Bioinform..

[26]  A. Tomasz,et al.  Inactivation of the srtA Gene Affects Localization of Surface Proteins and Decreases Adhesion of Streptococcus pneumoniae to Human Pharyngeal Cells In Vitro , 2003, Infection and Immunity.

[27]  R. Losick,et al.  Additional Targets of the Bacillus subtilis Global Regulator CodY Identified by Chromatin Immunoprecipitation and Genome-Wide Transcript Analysis , 2003, Journal of bacteriology.

[28]  K. Ochi,et al.  RelA Protein Is Involved in Induction of Genetic Competence in Certain Bacillus subtilis Strains by Moderating the Level of Intracellular GTP , 2002, Journal of bacteriology.

[29]  P. Andrew,et al.  Role of Inflammatory Mediators in Resistance and Susceptibility to Pneumococcal Infection , 2002, Infection and Immunity.

[30]  H. Malke,et al.  relA-Independent Amino Acid Starvation Response Network of Streptococcus pyogenes , 2001, Journal of bacteriology.

[31]  Jeremy S. Brown,et al.  Immunization with Components of Two Iron Uptake ABC Transporters Protects Mice against Systemic Streptococcus pneumoniae Infection , 2001, Infection and Immunity.

[32]  Elliot J. Lefkowitz,et al.  Genome of the Bacterium Streptococcus pneumoniae Strain R6 , 2001, Journal of bacteriology.

[33]  S. Salzberg,et al.  Complete Genome Sequence of a Virulent Isolate of Streptococcus pneumoniae , 2001, Science.

[34]  A D Long,et al.  Improved Statistical Inference from DNA Microarray Data Using Analysis of Variance and A Bayesian Statistical Framework , 2001, The Journal of Biological Chemistry.

[35]  P. Renault,et al.  Pleiotropic transcriptional repressor CodY senses the intracellular pool of branched‐chain amino acids in Lactococcus lactis , 2001, Molecular microbiology.

[36]  Jeremy S. Brown,et al.  A Streptococcus pneumoniae pathogenicity island encoding an ABC transporter involved in iron uptake and virulence , 2001, Molecular microbiology.

[37]  O. Kuipers Complete DNA sequence of Lactococcus lactis adds flavor to genomics. , 2001, Genome research.

[38]  S. Ehrlich,et al.  The complete genome sequence of the lactic acid bacterium Lactococcus lactis ssp. lactis IL1403. , 2001, Genome research.

[39]  H. Malke,et al.  Life in protein‐rich environments: the relA‐independent response of Streptococcus pyogenes to amino acid starvation , 2000, Molecular microbiology.

[40]  J. Claverys,et al.  Is the Ami-AliA/B oligopeptide permease of Streptococcus pneumoniae involved in sensing environmental conditions? , 2000, Research in microbiology.

[41]  Aras Kadioglu,et al.  Host Cellular Immune Response to Pneumococcal Lung Infection in Mice , 2000, Infection and Immunity.

[42]  C. J. Thomson,et al.  New Gene Cassettes for Trimethoprim Resistance,dfr13, and Streptomycin-Spectinomycin Resistance,aadA4, Inserted on a Class 1 Integron , 2000, Antimicrobial Agents and Chemotherapy.

[43]  J. García,et al.  Molecular characterization of PcpA: a novel choline-binding protein of Streptococcus pneumoniae. , 1998, FEMS microbiology letters.

[44]  P. Serror,et al.  CodY is required for nutritional repression of Bacillus subtilis genetic competence , 1996, Journal of bacteriology.

[45]  A. Sonenshein,et al.  Interaction of Cody, a novel Bacillus subtillis DNA‐binding protein, with the dpp promoter region , 1996, Molecular microbiology.

[46]  C. Rosenow,et al.  Pyruvate oxidase, as a determinant of virulence in Streptococcus pneumoniae , 1996, Molecular microbiology.

[47]  H. Masure,et al.  Peptide permeases from Streptococcus pneumoniae affect adherence to eucaryotic cells , 1995, Infection and immunity.

[48]  Jan Kok,et al.  MicroPreP: a cDNA microarray data pre-processing framework. , 2003, Applied bioinformatics.