TetR Family Member PsrA Directly Binds the Pseudomonas rpoS and psrA Promoters

ABSTRACT We have previously described a Pseudomonas gene, psrA, which enhances transcription of the rpoS sigma factor gene at stationary phase. We present molecular data which demonstrate that in Pseudomonas putida PsrA binds specifically to the rpoS and psrA promoters in DNA regions having similar palindromic sequences, C/GAAAC N2-4 GTTTG/C, where N is any nucleotide. The position of the initiation of transcription was determined for both promoters, and PsrA binds from positions −59 to −35 in the rpoS promoter and from −18 to +20 in the psrA promoter with respect to the +1 transcription site. Expression studies with a psrA-lacZ transcriptional fusion in wild-type and psrA::Tn5 knockout mutants revealed that psrA was under additional control in response to growth phase. A model for the role of PsrA in the regulation of rpoS and psrA is presented.

[1]  Herman P. Spaink,et al.  Promoters in the nodulation region of the Rhizobium leguminosarum Sym plasmid pRL1JI , 2004, Plant Molecular Biology.

[2]  U. K. Laemmli,et al.  Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4 , 1970, Nature.

[3]  D. Helinski,et al.  Replication of an origin-containing derivative of plasmid RK2 dependent on a plasmid function provided in trans. , 1979, Proceedings of the National Academy of Sciences of the United States of America.

[4]  R. Hengge-aronis,et al.  The cellular concentration of the sigma S subunit of RNA polymerase in Escherichia coli is controlled at the levels of transcription, translation, and protein stability. , 1994, Genes & development.

[5]  V. Venturi,et al.  Regulation of rpoS Gene Expression in Pseudomonas: Involvement of a TetR Family Regulator , 2001, Journal of bacteriology.

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

[7]  C. Cunning,et al.  Promoter Substitution and Deletion Analysis of Upstream Region Required for rpoS Translational Regulation , 1998, Journal of bacteriology.

[8]  Kan Tanaka,et al.  Transcription of the principal sigma‐factor genes, rpoD and rpoS, in Pseudomonas aeruginosa is controlled according to the growth phase , 1994, Molecular microbiology.

[9]  D. Hassett,et al.  Effect of rpoS Mutation on the Stress Response and Expression of Virulence Factors in Pseudomonas aeruginosa , 1999, Journal of bacteriology.

[10]  S. Ueda,et al.  Regulation of RNA polymerase sigma subunit synthesis in Escherichia coli: intracellular levels of four species of sigma subunit under various growth conditions , 1996, Journal of bacteriology.

[11]  C. Whistler,et al.  The Two-Component Regulators GacS and GacA Influence Accumulation of the Stationary-Phase Sigma Factor ςS and the Stress Response in Pseudomonas fluorescensPf-5 , 1998, Journal of bacteriology.

[12]  C. Yanisch-Perron,et al.  Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. , 1985, Gene.

[13]  E. Greenberg,et al.  Regulation of Quorum Sensing by RpoS inPseudomonas aeruginosa , 2000, Journal of bacteriology.

[14]  K. Tanaka,et al.  A hierarchical quorum‐sensing cascade in Pseudomonas aeruginosa links the transcriptional activators LasR and RhIR (VsmR) to expression of the stationary‐phase sigma factor RpoS , 1996, Molecular microbiology.

[15]  S. Gottesman,et al.  Signal Transduction Cascade for Regulation of RpoS: Temperature Regulation of DsrA , 2001, Journal of bacteriology.

[16]  M. Bally,et al.  RpoS-dependent stress tolerance in Pseudomonas aeruginosa. , 1999, Microbiology.

[17]  B. Schippers,et al.  Reduction of Yield Depressions in High Frequency Potato Cropping Soil after Seed Tuber Treatments with Antagonistic Fluorescent Pseudomonas spp. , 1983 .

[18]  J. Colmer,et al.  The Pseudomonas aeruginosa Exotoxin A Regulatory Gene, ptxS: Evidence for Negative Autoregulation , 1999, Journal of bacteriology.

[19]  R. Hengge-aronis,et al.  The role of the sigma factor sigma S (KatF) in bacterial global regulation. , 1994, Annual review of microbiology.

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

[21]  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.

[22]  T. Silhavy,et al.  The LysR Homolog LrhA Promotes RpoS Degradation by Modulating Activity of the Response Regulator SprE , 1999, Journal of bacteriology.