Protection from Visible Light Is Important for Burkholderia Glumae of Katg the Quorum Sensing-dependent Gene

ABSTRACT Quorum sensing (QS) plays important roles in the pathogenicity of Burkholderia glumae, the causative agent of bacterial rice grain rot. We determined how QS is involved in catalase expression in B. glumae. The QS-defective mutant of B. glumae exhibited less catalase activity than wild-type B. glumae. A β-glucuronidase assay of a katG::Tn3-gusA78 reporter fusion protein revealed that katG expression is under the control of QS. Furthermore, katG expression was upregulated by QsmR, a transcriptional activator for flagellar-gene expression that is regulated by QS. A gel mobility shift assay confirmed that QsmR directly activates katG expression. The katG mutant produced toxoflavin but exhibited less severe disease than BGR1 on rice panicles. Under visible light conditions and a photon flux density of 61.6 μmol−1 m−2, the survival rate of the katG mutant was 105-fold lower than that of BGR1. This suggests that KatG is a major catalase that protects bacterial cells from visible light, which probably results in less severe disease caused by the katG mutant.

[1]  Deog-Yong Lee,et al.  Identification of quorum sensing-related regulons in Vibrio vulnificus by two-dimensional gel electrophoresis and differentially displayed reverse transcriptase PCR. , 2007, FEMS immunology and medical microbiology.

[2]  K. Iiyama,et al.  A role of phytotoxin in virulence of Pseudomonas glumae Kurita et Tabei , 1995 .

[3]  Jaime L. Sajecki,et al.  Role of Catalase in Campylobacter jejuniIntracellular Survival , 2000, Infection and Immunity.

[4]  Hiroaki Suga,et al.  Quorum sensing and the LysR‐type transcriptional activator ToxR regulate toxoflavin biosynthesis and transport in Burkholderia glumae , 2004, Molecular microbiology.

[5]  M. Bevan,et al.  GUS fusions: beta‐glucuronidase as a sensitive and versatile gene fusion marker in higher plants. , 1987, The EMBO journal.

[6]  D. Hassett,et al.  Role of the Pseudomonas aeruginosa oxyR-recG Operon in Oxidative Stress Defense and DNA Repair: OxyR-Dependent Regulation of katB-ankB, ahpB, andahpC-ahpF , 2000, Journal of bacteriology.

[7]  S. Farr,et al.  Oxidative stress responses in Escherichia coli and Salmonella typhimurium. , 1991, Microbiological reviews.

[8]  P. Nicholls,et al.  Enzymology and structure of catalases , 2000 .

[9]  Kim Rutherford,et al.  Genomic plasticity of the causative agent of melioidosis, Burkholderia pseudomallei. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[10]  W. Berends,et al.  On the origin of the toxicity of toxoflavin. , 1961, Biochimica et biophysica acta.

[11]  S. Diggle,et al.  Quorum sensing regulates dpsA and the oxidative stress response in Burkholderia pseudomallei. , 2006, Microbiology.

[12]  B. Kan,et al.  Quorum Sensing Enhances the Stress Response in Vibrio cholerae , 2007, Applied and Environmental Microbiology.

[13]  P. Stewart,et al.  Quorum sensing in Pseudomonas aeruginosa controls expression of catalase and superoxide dismutase genes and mediates biofilm susceptibility to hydrogen peroxide , 1999, Molecular microbiology.

[14]  C. Napoli,et al.  Molecular characterization of cloned avirulence genes from race 0 and race 1 of Pseudomonas syringae pv. glycinea , 1987, Journal of bacteriology.

[15]  S. Pan,et al.  An Agrobacterium catalase is a virulence factor involved in tumorigenesis , 2000, Molecular microbiology.

[16]  Ingyu Hwang,et al.  Toxoflavin Produced by Burkholderia glumae Causing Rice Grain Rot Is Responsible for Inducing Bacterial Wilt in Many Field Crops. , 2003, Plant disease.

[17]  Minkyun Kim,et al.  Regulation of polar flagellum genes is mediated by quorum sensing and FlhDC in Burkholderia glumae , 2007, Molecular microbiology.

[18]  G. Storz,et al.  OxyR, a positive regulator of hydrogen peroxide-inducible genes in Escherichia coli and Salmonella typhimurium, is homologous to a family of bacterial regulatory proteins. , 1989, Proceedings of the National Academy of Sciences of the United States of America.

[19]  T. Laessig,et al.  Cu,Zn Superoxide Dismutase of Mycobacterium tuberculosis Contributes to Survival in Activated Macrophages That Are Generating an Oxidative Burst , 2001, Infection and Immunity.

[20]  Alex Levine,et al.  H2O2 from the oxidative burst orchestrates the plant hypersensitive disease resistance response , 1994, Cell.

[21]  G. Diaz,et al.  A double staining method for differentiating between two classes of mycobacterial catalase in polyacrylamide electrophoresis gels. , 1986, Analytical biochemistry.

[22]  S. Farr,et al.  Oxidative stress responses in Escherichia coli and Salmonella typhimurium , 1991 .

[23]  J. Imlay,et al.  Pathways of oxidative damage. , 2003, Annual review of microbiology.

[24]  Isaac Ginsburg,et al.  Mechanism of Visible Light Phototoxicity on Porphyromonas gingivalis and Fusobacterium nucleatum , 2005, Photochemistry and photobiology.

[25]  E. Greenberg,et al.  Quorum sensing in bacteria: the LuxR-LuxI family of cell density-responsive transcriptional regulators , 1994, Journal of bacteriology.

[26]  D. Hassett,et al.  Cloning and characterization of the katB gene of Pseudomonas aeruginosa encoding a hydrogen peroxide-inducible catalase: purification of KatB, cellular localization, and demonstration that it is essential for optimal resistance to hydrogen peroxide , 1995, Journal of bacteriology.

[27]  S. Mongkolsuk,et al.  A Xanthomonas Alkyl Hydroperoxide Reductase Subunit C (ahpC) Mutant Showed an Altered Peroxide Stress Response and Complex Regulation of the Compensatory Response of Peroxide Detoxification Enzymes , 2000, Journal of bacteriology.

[28]  G. Martin,et al.  Strategies used by bacterial pathogens to suppress plant defenses. , 2004, Current opinion in plant biology.

[29]  M. Berenbaum,et al.  SANGUINARINE, A PHOTOTOXIC H2O2‐PRODUCING ALKALOID , 1989, Photochemistry and photobiology.

[30]  S. Mongkolsuk,et al.  Regulation of the katG‐dpsA operon and the importance of KatG in survival of Burkholderia pseudomallei exposed to oxidative stress , 2003, FEBS letters.

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

[32]  S. Mongkolsuk,et al.  The Burkholderia pseudomallei oxyR gene: expression analysis and mutant characterization. , 2002, Gene.

[33]  D. Hassett,et al.  A Protease-Resistant Catalase, KatA, Released upon Cell Lysis during Stationary Phase Is Essential for Aerobic Survival of a Pseudomonas aeruginosa oxyR Mutant at Low Cell Densities , 2000, Journal of bacteriology.

[34]  M. Swanson,et al.  Legionella pneumophila Catalase-Peroxidases Are Required for Proper Trafficking and Growth in Primary Macrophages , 2003, Infection and Immunity.