Transcriptional regulation under pressure conditions by RNA polymerase σ54 factor with a two-component regulatory system in Shewanella violacea

Abstract. Deep-sea bacteria have unique systems for gene and protein expression controlled by hydrostatic pressure. One of the σ factors, σ54, was found to play an important role in pressure-regulated transcription in a deep-sea piezophilic bacterium, Shewanella violacea. A glutamine synthetase gene (glnA) has been targeted as a model for the pressure-regulated promoter to investigate transcriptional regulation by the σ54 factor. Recognition sites for σ54 and σ70 factors were observed at an upstream region of the glnA, and NtrC-binding sites were also identified at the same region. Primer extension analyses revealed that the transcription initiation sites of both promoters were determined and that transcription from the σ54 site was regulated by elevated pressure. The σ54 promoter is known to be activated by a two-component signal transduction system, the NtrB–NtrC phosphorylation relay. Our results suggested that this system might be regulated by deep-sea conditions and that the gene expression controlled by the σ54 promoter was actually regulated by pressure. We propose a possible model of the molecular mechanisms for pressure-regulated transcription.

[1]  K. Horikoshi,et al.  Isolation and properties of barophilic and barotolerant bacteria from deep-sea mud samples , 1995, Biodiversity & Conservation.

[2]  M. Merrick,et al.  Positive and negative control of the glnA ntrBC regulon in Klebsiella pneumoniae. , 1984, EMBO Journal.

[3]  W. C. Bowman,et al.  In Vitro Reconstitution and Characterization of the Rhodobacter capsulatus NtrB and NtrC Two-component System (*) , 1996, The Journal of Biological Chemistry.

[4]  K. Horikoshi,et al.  Structural Analysis of the ntrBC Genes of Deep-sea Piezophilic Shewanella violacea , 2000, Bioscience, biotechnology, and biochemistry.

[5]  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.

[6]  K. Horikoshi,et al.  Isolation and piezoresponse of the rpoA gene encoding the RNA polymerase alpha subunit from the deep-sea piezophilic bacterium Shewanella violacea. , 2000, FEMS microbiology letters.

[7]  A. Ninfa,et al.  Initiation of transcription at the bacterial glnAp2 promoter by purified E. coli components is facilitated by enhancers , 1987, Cell.

[8]  S. Sasse-Dwight,et al.  Role of eukaryotic-type functional domains found in the prokaryotic enhancer receptor factor σ 54 , 1990, Cell.

[9]  B. Magasanik,et al.  Transcription of glnA by purified Escherichia coli components: core RNA polymerase and the products of glnF, glnG, and glnL. , 1985, Proceedings of the National Academy of Sciences of the United States of America.

[10]  C. Gross,et al.  Consensus sequence for Escherichia coli heat shock gene promoters. , 1985, Proceedings of the National Academy of Sciences of the United States of America.

[11]  S. Harayama,et al.  Involvement of Pseudomonas putida RpoN sigma factor in regulation of various metabolic functions , 1989, Journal of bacteriology.

[12]  B. Magasanik,et al.  Transcription of glnA in E. coli is stimulated by activator bound to sites far from the promoter , 1986, Cell.

[13]  David J. Studholme,et al.  The Bacterial Enhancer-Dependent ς54(ςN) Transcription Factor , 2000, Journal of bacteriology.

[14]  K. Horikoshi,et al.  Cloning and characterization of the gene encoding RNA polymerase sigma factor sigma(54) of deep-sea piezophilic Shewanella violacea. , 2000, Biochimica et biophysica acta.

[15]  Y. Nakamura,et al.  Glutamine synthetase gene expression at elevated hydrostatic pressure in a deep-sea piezophilic Shewanella violacea. , 2000, FEMS microbiology letters.

[16]  K. Horikoshi,et al.  Mechanisms of gene expression controlled by pressure in deep-sea microorganisms , 1998, Extremophiles.

[17]  R. Edwards,et al.  Nitrogen control in bacteria. , 1995, Microbiological reviews.

[18]  A. Böck,et al.  Characterization of divergent NtrA‐dependent promoters in the anaerobically expressed gene cluster coding for hydrogenase 3 components of Escherichia coli , 1990, Molecular microbiology.

[19]  R. Dixon Tandem promoters determine regulation of the Klebsiella pneumoniae glutamine synthetase (glnA) gene. , 1984, Nucleic acids research.

[20]  S. Kustu,et al.  Protein kinase and phosphoprotein phosphatase activities of nitrogen regulatory proteins NTRB and NTRC of enteric bacteria: roles of the conserved amino-terminal domain of NTRC. , 1988, Proceedings of the National Academy of Sciences of the United States of America.

[21]  K. Horikoshi,et al.  Structure of genes in a pressure-regulated operon and adjacent regions from a barotolerant bacterium strain DSS12 , 1997 .