Kukavica-IbruljSanschagrinPetersonEtAl2008

Référence

Kukavica-Ibrulj, I., Sanschagrin, F., Peterson, A., Whiteley, M., Boyle, B., MacKay, J., Levesque, R.C. (2008) Functional genomics of PycR, a LysR family transcriptional regulator essential for maintenance of Pseudomonas aeruginosa in the rat lung. Microbiology-Sgm, 154:2106-2118.

Résumé

The human opportunistic pathogen Pseudomonas aeruginosa is the major cause of morbidity and mortality of cystic fibrosis patients and is responsible for a variety of infections in compromised hosts. Using PCR-based signature-tagged mutagenesis, we identified a P. aeruginosa STM5437 mutant with an insertion into the PA5437 gene (called pycR for putative pyruvate carboxylase regulator). PycR inactivation results in 100 000-fold attenuation of virulence in the rat lung in vivo. PycR has the signature of a transcriptional regulator with a predicted helix-turn-helix motif binding to a typical LysR DNA binding site in the PA5436 (pycA)-PA5437 (pycR) intercistronic region. Two pyruvate carboxylase subunits (pycA and pycB) are divergently transcribed upstream of pycR. Transcriptional start sites of pycR and pycA are located at -127 and -88 bp upstream of their initiation codons with Shine-Dalgarno and putative promoter sequences containing -10 and -35 sequences. The DNA binding of PycR was confirmed by DNA mobility shift assay. Genome-wide transcriptional profiling and quantitative real-time PCR (qRT-PCR) indicated that the genes differentially regulated by PycR include two pyruvate carboxylase genes and genes necessary for lipid metabolism, lipolytic activity, anaerobic respiration and biofilm formation. PycR is a regulator with pleiotropic effects on virulence factors, such as lipase and esterase expression and biofilm formation, which are important for maintenance of P. aeruginosa in chronic lung infection.

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@ARTICLE { Kukavica-IbruljSanschagrinPetersonEtAl2008,
    AUTHOR = { Kukavica-Ibrulj, I. and Sanschagrin, F. and Peterson, A. and Whiteley, M. and Boyle, B. and MacKay, J. and Levesque, R.C. },
    TITLE = { Functional genomics of PycR, a LysR family transcriptional regulator essential for maintenance of Pseudomonas aeruginosa in the rat lung },
    JOURNAL = { Microbiology-Sgm },
    YEAR = { 2008 },
    VOLUME = { 154 },
    PAGES = { 2106-2118 },
    MONTH = { jul },
    AF = { Kukavica-Ibrulj, IrenaEOLEOLSanschagrin, FrancoisEOLEOLPeterson, AshleyEOLEOLWhiteley, MarvinEOLEOLBoyle, BrianEOLEOLMacKay, JohnEOLEOLLevesque, Roger C. },
    DI = { 10.1099/mic.0.2007/011239-0 },
    PG = { 13 },
    PN = { Part 7 },
    SN = { 1350-0872 },
    UT = { ISI:000257893800026 },
    ABSTRACT = { The human opportunistic pathogen Pseudomonas aeruginosa is the major cause of morbidity and mortality of cystic fibrosis patients and is responsible for a variety of infections in compromised hosts. Using PCR-based signature-tagged mutagenesis, we identified a P. aeruginosa STM5437 mutant with an insertion into the PA5437 gene (called pycR for putative pyruvate carboxylase regulator). PycR inactivation results in 100 000-fold attenuation of virulence in the rat lung in vivo. PycR has the signature of a transcriptional regulator with a predicted helix-turn-helix motif binding to a typical LysR DNA binding site in the PA5436 (pycA)-PA5437 (pycR) intercistronic region. Two pyruvate carboxylase subunits (pycA and pycB) are divergently transcribed upstream of pycR. Transcriptional start sites of pycR and pycA are located at -127 and -88 bp upstream of their initiation codons with Shine-Dalgarno and putative promoter sequences containing -10 and -35 sequences. The DNA binding of PycR was confirmed by DNA mobility shift assay. Genome-wide transcriptional profiling and quantitative real-time PCR (qRT-PCR) indicated that the genes differentially regulated by PycR include two pyruvate carboxylase genes and genes necessary for lipid metabolism, lipolytic activity, anaerobic respiration and biofilm formation. PycR is a regulator with pleiotropic effects on virulence factors, such as lipase and esterase expression and biofilm formation, which are important for maintenance of P. aeruginosa in chronic lung infection. },
    KEYWORDS = { SIGNATURE-TAGGED MUTAGENESIS; YEAST PYRUVATE-CARBOXYLASE; GENE-EXPRESSION; MICROARRAY ANALYSIS; TWITCHING MOTILITY; MOLECULAR-BIOLOGY; BIOFILM FORMATION; IDENTIFICATION; SEQUENCE; SYSTEM },
    OWNER = { brugerolles },
    TIMESTAMP = { 2008.08.18 },
}

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