Differential pressure resistance in the activity of RNA polymerase isolated from Shewanella violacea and Escherichia coli

RNA polymerase was purified from the piezophile Shewanella violacea DSS12, and the transcriptional activity after pressure treatment was compared with that of the mesophile Escherichia coli. Application of pressure at 100 MPa for 30 min reduced the E. coli RNA polymerase activity to 60% of the activity at atmospheric pressure, whereas the S. violacea RNA polymerase maintained full activity, indicating that the S. violacea RNA polymerase is more stable than its E. coli counterpart. This result was supported by the analysis of the strength of subunit interactions of the enzyme from both species, using a high-pressure electrophoresis apparatus, which showed that a pressure of 140 MPa caused dissociation of E. coli RNA polymerase but not that of S. violacea RNA polymerase. On the other hand, the core enzyme of S. violacea RNA polymerase, which lacked the σ70 factor, was dissociated at 140 MPa. These results suggest that the σ70 factor is required for stabilization of S. violacea RNA polymerase under high-pressure conditions. In this paper, we provide in vitro evidence for piezoadaptation at the transcriptional level, using purified RNA polymerase from cells of S. violacea and E. coli.

[1]  D. Bartlett.  Pressure effects on in vivo microbial processes. , 2002, Biochimica et biophysica acta.

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

[3]  D. Chatterji,et al.  An improved method for the purification of DNA-dependent RNA polymerase from Escherichia coli. , 1988, Journal of Biochemical and Biophysical Methods.

[4]  A. Gunasekera,et al.  Binding of the sigma 70 protein to the core subunits of Escherichia coli RNA polymerase, studied by iron-EDTA protein footprinting. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

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

[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. Ishihama,et al.  Genetics of bacterial RNA polymerases. , 1979, Annual review of genetics.

[8]  D. Bartlett,et al.  Isolation and characterization of the gene encoding single-stranded-DNA-binding protein (SSB) from four marine Shewanella strains that differ in their temperature and pressure optima for growth. , 1997, Microbiology.

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

[10]  Jerson L. Silva,et al.  Pressure stability of proteins. , 1993, Annual review of physical chemistry.

[11]  O. White,et al.  Genome sequence of the dissimilatory metal ion–reducing bacterium Shewanella oneidensis , 2002, Nature Biotechnology.

[12]  K. Horikoshi,et al.  Pressure-regulated metabolism in microorganisms , 1999 .

[13]  Chiaki Kato,et al.  Transcriptional regulation under pressure conditions by RNA polymerase σ54 factor with a two-component regulatory system in Shewanella violacea , 2002, Extremophiles.

[14]  R. Clegg,et al.  Heterogeneity of E. coli RNA polymerase revealed by high pressure. , 1995, Journal of molecular biology.

[15]  R. Burgess,et al.  Use of Mono Q high-resolution ion-exchange chromatography to obtain highly pure and active Escherichia coli RNA polymerase. , 1990, Biochemistry.

[16]  D. Bartlett,et al.  Comparison of high pressure-induced dissociation of single-stranded DNA-binding protein (SSB) from high pressure-sensitive and high pressure-adapted marine Shewanella species , 2002, Extremophiles.

[17]  M. Gross,et al.  Proteins under pressure. The influence of high hydrostatic pressure on structure, function and assembly of proteins and protein complexes. , 1994, European journal of biochemistry.

[18]  M. Fujita,et al.  Purification and characterization of a DNA-dependent RNA polymerase from Pseudomonas putida. , 1992, Bioscience, biotechnology, and biochemistry.

[19]  T. Welch,et al.  Identification of a regulatory protein required for pressure‐responsive gene expression in the deep‐sea bacterium Photobacterium species strain SS9 , 1998, Molecular microbiology.

[20]  K. Horikoshi,et al.  Purification and characterization of RNA polymerase from deep-sea piezophilic Shewanella violacea , 2002 .

[21]  K. Horikoshi,et al.  Taxonomic studies of deep-sea barophilic Shewanella strains and description of Shewanella violacea sp. nov. , 1998, Archives of Microbiology.

[22]  R. Burgess,et al.  A procedure for the rapid, large-scall purification of Escherichia coli DNA-dependent RNA polymerase involving Polymin P precipitation and DNA-cellulose chromatography. , 1975, Biochemistry.

[23]  R. Clegg,et al.  High pressure electrophoresis in narrow bore glass tubes: One‐ and two‐dimensional separations of protein subunits , 1996 .

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

[25]  Rainer Jaenicke,et al.  Proteins under pressure , 1994 .

[26]  M. Silverman,et al.  Isolation of a gene regulated by hydrostatic pressure in a deep-sea bacterium , 1989, Nature.

[27]  C. Wu,et al.  Conformational transition of Escherichia coli RNA polymerase induced by the interaction of sigma subunit with core enzyme. , 1976, Biochemistry.

[28]  S. Darst,et al.  Crystal Structure of a σ70 Subunit Fragment from E. coli RNA Polymerase , 1996, Cell.