The PerR regulon in peroxide resistance and virulence of Streptococcus pyogenes

Prior studies have shown that the catalase‐deficient pathogen Streptococcus pyogenes (group A streptococcus) has a robust ability to resist oxidative stress that partially involves the transcriptional regulator PerR. However, the extent of the PerR regulon and the contribution of the members of this regulon to virulence are unknown. In this study, DNase I footprinting revealed that PerR binds specifically to a single site upstream of the promoter for the gene encoding alkyl hydroperoxide reductase (ahpC). However, analyses of transcript abundance revealed that while ahpC is regulated in response to growth phase, its regulation is independent of PerR. Instead, PerR regulates transcription of a divergent gene cluster that encodes a putative cold shock protein. The gene encoding the Dps‐like peroxide resistance protein MrgA was repressed by PerR, consistent with the presence of a PerR binding site in its promoter. Phenotypic analyses of PerR–, AhpC– and MrgA– mutants revealed that while AhpC is not essential for resistance to challenge with hydrogen peroxide in vitro, AhpC does contribute to scavenging of endogenous hydrogen peroxide and is required for virulence in a murine model of infection. In contrast, a MrgA– mutant was hypersensitive to challenge with peroxide in vitro, but was fully virulent in all animal models tested. Finally, a PerR– mutant was hyper‐resistant to peroxide, yet was highly attenuated for virulence in all murine models. These data demonstrate that while a mutant's capacity to resist peroxide stress did not directly correlate with its ability to cause disease, the appropriate regulation of the peroxide stress response is critical for virulence.

[1]  M. Caparon,et al.  Contribution of Glutathione Peroxidase to the Virulence of Streptococcus pyogenes , 2004, Infection and Immunity.

[2]  S. Haataja,et al.  Molecular Basis of H2O2 Resistance Mediated by Streptococcal Dpr , 2003, The Journal of Biological Chemistry.

[3]  P. Fawcett,et al.  The Global Transcriptional Response of Bacillus subtilis to Peroxide Stress Is Coordinated by Three Transcription Factors , 2003, Journal of bacteriology.

[4]  J. Helmann,et al.  Recognition of DNA by Fur: a Reinterpretation of the Fur Box Consensus Sequence , 2002, Journal of bacteriology.

[5]  L. Björck,et al.  The Regulator PerR Is Involved in Oxidative Stress Response and Iron Homeostasis and Is Necessary for Full Virulence of Streptococcus pyogenes , 2002, Infection and Immunity.

[6]  J. Helmann,et al.  A peroxide‐induced zinc uptake system plays an important role in protection against oxidative stress in Bacillus subtilis , 2002, Molecular microbiology.

[7]  J. Pfeifer,et al.  Streptococcus-Zebrafish Model of Bacterial Pathogenesis , 2002, Infection and Immunity.

[8]  J. Helmann,et al.  Regulation of the Bacillus subtilis fur and perR Genes by PerR: Not All Members of the PerR Regulon Are Peroxide Inducible , 2002, Journal of bacteriology.

[9]  Y. Kamio,et al.  An Iron-Binding Protein, Dpr, from Streptococcus mutans Prevents Iron-Dependent Hydroxyl Radical Formation In Vitro , 2002, Journal of bacteriology.

[10]  J. Imlay,et al.  Alkyl Hydroperoxide Reductase Is the Primary Scavenger of Endogenous Hydrogen Peroxide in Escherichia coli , 2001, Journal of bacteriology.

[11]  J. Helmann,et al.  Roles of metal ions and hydrogen peroxide in modulating the interaction of the Bacillus subtilis PerR peroxide regulon repressor with operator DNA , 2001, Molecular microbiology.

[12]  S. Foster,et al.  PerR Controls Oxidative Stress Resistance and Iron Storage Proteins and Is Required for Virulence in Staphylococcus aureus , 2001, Infection and Immunity.

[13]  Bruce A. Roe,et al.  Complete genome sequence of an M1 strain of Streptococcus pyogenes , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[14]  B. Limbago,et al.  Role of Streptolysin O in a Mouse Model of Invasive Group A Streptococcal Disease , 2000, Infection and Immunity.

[15]  M. Caparon,et al.  Aerotolerance and Peroxide Resistance in Peroxidase and PerR Mutants of Streptococcus pyogenes , 2000, Journal of bacteriology.

[16]  Patrick Griffin,et al.  Peroxynitrite reductase activity of bacterial peroxiredoxins , 2000, Nature.

[17]  D. Heinrichs,et al.  Identification and Characterization of a Membrane Permease Involved in Iron-Hydroxamate Transport inStaphylococcus aureus , 2000, Journal of bacteriology.

[18]  Y. Kamio,et al.  Role of the dpr Product in Oxygen Tolerance in Streptococcus mutans , 2000, Journal of bacteriology.

[19]  M. Cunningham,et al.  Pathogenesis of group A streptococcal infections. , 2000, Clinical microbiology reviews.

[20]  D. Parsonage,et al.  The RofA Binding Site in Streptococcus pyogenes Is Utilized in Multiple Transcriptional Pathways , 2000, Journal of bacteriology.

[21]  M. Caparon,et al.  Contribution of NADH Oxidase to Aerobic Metabolism of Streptococcus pyogenes , 2000, Journal of bacteriology.

[22]  D. Low,et al.  Assessment of bacterial pathogenesis by analysis of gene expression in the host. , 2000, Annual review of genetics.

[23]  S Rozen,et al.  Primer3 on the WWW for general users and for biologist programmers. , 2000, Methods in molecular biology.

[24]  M. Stinson,et al.  Influence of Environmental Conditions on Hydrogen Peroxide Formation by Streptococcus gordonii , 1999, Infection and Immunity.

[25]  J. Musser,et al.  Extracellular Cysteine Protease Produced by Streptococcus pyogenes Participates in the Pathogenesis of Invasive Skin Infection and Dissemination in Mice , 1999, Infection and Immunity.

[26]  D. Hendrixson,et al.  The Haemophilus influenzae Hap serine protease promotes adherence and microcolony formation, potentiated by a soluble host protein. , 1998, Molecular cell.

[27]  J. Helmann,et al.  Bacillus subtilis contains multiple Fur homologues: identification of the iron uptake (Fur) and peroxide regulon (PerR) repressors , 1998, Molecular microbiology.

[28]  R. Kolter,et al.  The crystal structure of Dps, a ferritin homolog that binds and protects DNA , 1998, Nature Structural Biology.

[29]  J. Miller,et al.  In vivo and ex vivo regulation of bacterial virulence gene expression. , 1998, Current opinion in microbiology.

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

[31]  S. Hollingshead,et al.  Role of putative virulence factors of Streptococcus pyogenes in mouse models of long-term throat colonization and pneumonia , 1997, Infection and immunity.

[32]  D. Guiney.  Regulation of bacterial virulence gene expression by the host environment. , 1997, The Journal of clinical investigation.

[33]  J. Helmann,et al.  Mutation of the Bacillus subtilis alkyl hydroperoxide reductase (ahpCF) operon reveals compensatory interactions among hydrogen peroxide stress genes , 1996, Journal of bacteriology.

[34]  J. Rheinwald,et al.  Hyaluronic acid capsule and the role of streptococcal entry into keratinocytes in invasive skin infection. , 1996, The Journal of clinical investigation.

[35]  M. Caparon,et al.  Insertional inactivation of Streptococcus pyogenes sod suggests that prtF is regulated in response to a superoxide signal , 1996, Journal of bacteriology.

[36]  P. Cleary,et al.  C5a peptidase alters clearance and trafficking of group A streptococci by infected mice , 1996, Infection and immunity.

[37]  J. Helmann,et al.  Bacillus subtilis MrgA is a Dps(PexB) homologue: evidence for metalloregulation of an oxidative‐stress gene , 1995, Molecular microbiology.

[38]  Jeff F. Miller,et al.  Ectopic expression of the flagellar regulon alters development of the bordetella-host interaction , 1995, Cell.

[39]  V. Fischetti,et al.  A method to isolate RNA from gram-positive bacteria and mycobacteria. , 1994, Analytical biochemistry.

[40]  M. Caparon,et al.  The identification of rofA, a positive‐acting regulatory component of prtF expression: use of an mγδ‐based shuttle mutagenesis strategy in Streptococcus pyogenes , 1994 .

[41]  E. Hanski,et al.  Adherence and fibronectin binding are environmentally regulated in the group A streptococci , 1993, Molecular microbiology.

[42]  E. Maguin,et al.  An M protein with a single C repeat prevents phagocytosis of Streptococcus pyogenes: use of a temperature‐sensitive shuttle vector to deliver homologous sequences to the chromosome of S. pyogenes , 1993, Molecular microbiology.

[43]  M. Caparon,et al.  Expression of protein F, the fibronectin-binding protein of Streptococcus pyogenes JRS4, in heterologous streptococcal and enterococcal strains promotes their adherence to respiratory epithelial cells , 1992, Infection and immunity.

[44]  M. Gilmore,et al.  PCR amplification of streptococcal DNA using crude cell lysates. , 1992, FEMS microbiology letters.

[45]  J. Musser,et al.  Murine model of cutaneous infection with gram-positive cocci , 1992, Infection and immunity.

[46]  M. Caparon,et al.  Genetic manipulation of pathogenic streptococci. , 1991, Methods in enzymology.

[47]  E. Kinney.  Primer of Biostatistics , 1987 .

[48]  H. Müller.  Detection of hydrogen peroxide produced by microorganisms on an ABTS peroxidase medium. , 1985, Zentralblatt fur Bakteriologie, Mikrobiologie, und Hygiene. Series A, Medical microbiology, infectious diseases, virology, parasitology.

[49]  W. Gilbert,et al.  Sequencing end-labeled DNA with base-specific chemical cleavages. , 1980, Methods in enzymology.

[50]  J. R. Scott,et al.  A turbid plaque-forming mutant of phage P1 that cannot lysogenize Escherichia coli. , 1974, Virology.

[51]  Molecular Basis of H 2 O 2 Resistance Mediated by Streptococcal Dpr DEMONSTRATION OF THE FUNCTIONAL INVOLVEMENT OF THE PUTATIVE FERROXIDASE CENTER BY SITE-DIRECTED MUTAGENESIS IN STREPTOCOCCUS SUIS * , 2022 .