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Molecular cloning of ompRS, a regulatory locus controlling production of outer membrane proteins in Erwinia carotovora subsp. carotovora

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Summary

A locus, ompRS, controlling synthesis of outer membrane proteins was cloned from Erwinia carotovora subsp. carotovora (Ecc) by complementation of an Escherichia coli ompR—envZ mutant. The Ecc ompRS locus was both structurally and functionally similar to the ompR—envZ operon controlling porin gene expression in E. coli as shown by DNA hybridization and complementation of E. coli ompR and envZ mutants. Furthermore, introduction of ompRS into E. coli Δ (ompR—envZ) strains restored the osmoregulation of the major outer membrane protein genes ompC and ompF Maxicell analysis of ompRS-carrying plasmids suggested that proteins similar in size to the E. coli ompR and envZ gene products were encoded by the Ecc ompR and ompS genes, respectively. Introduction of an ompRS transposon mutant onto the Ecc chromosome by marker exchange mutagenesis showed that ompRS is essential for production of a major outer membrane porin in Ecc. This mutational defect could be complemented by clones carrying Ecc ompRS or E. coli ompR envZ. The lack of the porin did not, however, compromise the virulence of these Ecc mutants.

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References

  • Bassford PJ, Diedrich DL, Schnaitman CA, Reeves P (1977) Outer membrane proteins of Escherichia coli. VI. Protein alterations in bacteriophage-resistant mutants. J Bacteriol 131:608–622

    Google Scholar 

  • Birnboim HC, Doly J (1979) A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res 7:1513–1523

    CAS  PubMed  Google Scholar 

  • Boyer HW, Roulland-Dussoix D (1969) A complementation analysis of the restriction and modification of DNA in Escherichia coli. J Mol Biol 41:459–472

    Google Scholar 

  • Casadaban MJ (1976) Transposition and fusion of the lac genes to selected promotors in Escherichia coli using bacteriophage lambda and Mu. J Mol Biol 104:541–555

    Google Scholar 

  • Carter P, Bedouelle H, Winter G (1985) Improved oligonucleotide site-directed mutagenesis using M13 vectors. Nucleic Acids Res 13:4431–4443

    Google Scholar 

  • Collmer A, Keen NT (1986) The role of pectic enzymes in plant pathogenesis. Annu Rev Phytopathol 24:383–409

    Google Scholar 

  • Dorman CJ, Chatfield S, Higgins CF, Hayward C, Dougan G (1989) Characterization of porin and ompR mutants of a virulent strain of Salmonella typhimurium: ompR mutants are attenuated in vivo. Infect Immun 57:2136–2140

    Google Scholar 

  • Expert D, Toussaint A (1985) Bacteriocin-resistant mutants of Erwinia chrysanthemi: possible involvement of iron acquisition in phytopathogenicity. J Bacteriol 163:221–227

    Google Scholar 

  • Feiss M, Siegele DA, Rudolph CF, Frackman S (1982) Cosmid DNA packaging in vivo. Gene 17:123–130

    Google Scholar 

  • Garrett S, Taylor RK, Silhavy TJ (1983) Isolation and characterization of chain-terminating nonsense mutations in a porin regulatory gene, envZ. J Bacteriol 156:62–69

    Google Scholar 

  • Hall MN, Silhavy TJ (1979) Transcriptional regulation of Escherichia coli K-12 major outer membrane protein 1 B. J Bacteriol 140:342–350

    Google Scholar 

  • Hall MN, Silhavy TJ (1981) The ompB locus and the regulation of the major outer membrane porin proteins of Escherichia coli. J Mol Biol 146:23–43

    Google Scholar 

  • Harkki A, Palva ET (1985) A lamB expression plasmid for extending the host range of phage to other enterobacteria. FEMS Microbiol Lett 27;183–187

    Google Scholar 

  • Hinton JCD, Sidebotham JM, Hyman LJ, Perombelon MCM, Salmond GPC (1989) Isolation and characterization of transposon-induced mutants of Erwinia carotovora subsp. atroseptica exhibiting reduced virulence. Mol Gen Genet 217:141–148

    Google Scholar 

  • Igo MM, Ninfa AJ, Silhavy TJ (1989) A bacterial environmental sensor that functions as a protein kinase and stimulates transcriptional activation. Genes Dev 3:598–605

    Google Scholar 

  • Igo MM, Slauch JM, Silhavy TJ (1990) Signal transduction in bacteria: kinases that control gene expression. New Biologist 2:5–9

    Google Scholar 

  • Kotoujansky A (1987) Molecular genetics of pathogenesis by softrot erwinias. Annu Rev Phytopathol 25:405–430

    Google Scholar 

  • Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature New Biol 227:680–685

    Google Scholar 

  • Leary JV, Fulbright DW (1982) Chromosomal genetics of Pseudomonas spp. and Erwinia spp. In: Mount MS, Lacy GH (eds) Phytopathogenic procaryotes, vol 2. Academic Press, New York London, pp 229–253

    Google Scholar 

  • Liljeström P (1986) The EnvZ protein of Salmonella typhimurium LT-2 and Escherichia coli K-12 is located in the cytoplasmic membrane. FEMS Microbiol Lett 36:145–150

    Google Scholar 

  • Liljeström P, Määttänen PL, Palva ET (1982) Cloning of the regulatory locus ompB of Salmonella typhimurium LT-2. II. Identification of the envZ gene product, a protein involved in the expression of porin proteins. Mol Gen Genet 188:190–194

    Google Scholar 

  • Liljeström P, Pirhonen M, Palva ET (1985) In vivo transfer of chromosomal mutations onto multicopy plasmids by transduction with bacteriophage P1. Gene 40:241–246

    Google Scholar 

  • Liljeström P, Laamanen 1, Palva ET (1988) Structure and expression of the ompB operon, the regulatory locus for the outer membrane porin regulon in Salmonella typhimurium LT-2. J Mol Biol 201:663–673

    Google Scholar 

  • Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning. A laboratory manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York

    Google Scholar 

  • Miller JH (1972) Experiments in molecular genetics. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York

    Google Scholar 

  • Palva ET (1978) Major outer membrane protein in Salmonella typhimurium induced by maltose. J Bacteriol 136:286–294

    CAS  PubMed  Google Scholar 

  • Palva ET (1979) Relationship between ompB genes of Escherichia coli and Salmonella typhimurium. FEMS Microbiol Lett 5:205–209

    Google Scholar 

  • Palva ET, Harkki A, Karkku H, Lång H, Pirhonen M (1987) λ vehicles provide new tools for genetic analysis of Gram-negative bacteria. Microbial Pathogenesis 3:227–230

    Google Scholar 

  • Pérombelon MCM, Kelman A (1980) Ecology of the soft rot erwinias. Annu Rev Phytopathol 18:361–387

    Google Scholar 

  • Pirhonen M, Palva ET (1988) Occurrence of bacteriophage T4 receptor in Erwinia carotovora. Mol Gen Genet 214:170–172

    Google Scholar 

  • Pirhonen M, Heino P, Helander I, Harju P, Palva ET (1988) Bacteriophage T4 resistant mutants of the plant pathogen Erwinia carotovora. Microbial Pathogenesis 4:359–367

    Google Scholar 

  • Roeder DL, Colimer A (1985) Marker-exchange mutagenesis of a pectate lyase isozyme gene in Erwinia chrysanthemi. J Bacteriol 164:51–56

    Google Scholar 

  • Ronson CW, Nixon BT, Ausubel FM (1987) Conserved domains in bacterial regulatory proteins that respond to environmental stimuli. Cell 49:579–581

    Google Scholar 

  • Saarilahti HT, Palva ET (1986) Major outer membrane proteins in the phytopathogenic bacteria Erwinia carotovora subsp. carotovora and subsp. atroseptica. FEMS Microbiol Lett 35:267–270

    Google Scholar 

  • Saarilahti HT, Palva ET (1990) Analysis of OmpC-β-lactamase hybrid proteins: OmpC appears not to contain discrete localization signals. FEMS Microbiol Lett 69:49–54

    Google Scholar 

  • Sancar A, Hack AM, Rupp WD (1979) Simple method for identification of plasmid coded proteins. J Bacteriol 137:692–693

    Google Scholar 

  • Schoonejans E, Expert D, Touissaint A (1987) Characterization and virulence properties of Erwinia chrysanthemi lipopolysaccharide-defective, φ EC2-resistantmutants. J Bacteriol 169:4011–4017

    Google Scholar 

  • Silhavy TJ, Berman ML, Enquist LW (1984) Experiments with gene fusions. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York

    Google Scholar 

  • Stock JB, Stock AM, Mottonen JM (1990) Signal transduction in bacteria. Nature 344:395–400

    Google Scholar 

  • Taylor RK, Hall MN, Enquist LW, Silhavy TJ (1981) Identification of OmpR: a positive regulatory protein controlling expression of the major outer membrane porin proteins of E. coli K-12. J Bacteriol 147:255–258

    Google Scholar 

  • Taylor RK, Hall MN, Silhavy TJ (1983) Isolation and characterization of mutations altering expression of the major outer membrane porin proteins using the local anesthetic procaine. J Mol Biol 166:273–282

    Google Scholar 

  • Torriani A (1960) Influence of inorganic phosphate in the formation of phosphatases by Escherichia coli. Biochim Biophys Acta 38:460–479

    Google Scholar 

  • van Alphen W, Lugtenberg B (1977) Influence of osmolarity of the growth medium on the outer membrane protein pattern of Escherichia coli. J Bacteriol 131:623–630

    Google Scholar 

  • Way JC, Davis MA, Morisato D; Roberts DE, Kleckner N (1984) New Tn10 derivatives for transposon mutagenesis and for construction of lacZ fusions by transposition. Gene 32:369–379

    Google Scholar 

  • Wilson GG, Young KKY, Edlin GJ, Konigsberg W (1979) High-frequency generalized transduction by bacteriophage T4. Nature 280:80–82

    Google Scholar 

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Author notes

  1. Hannu T. Saarilahti

    Present address: National Public Health Service, Mannerheimintie 166, SF-00300, Helsinki, Finland

Authors and Affiliations

  1. Department of Molecular Genetics, Uppsala Genetic Center, Swedish University of Agricultural Sciences, Box 7003, S-75007, Uppsala, Sweden

    Maj-Britt Karlsson, Minna Pirhonen, Hannu T. Saarilahti & E. Tapio Palva

Authors
  1. Maj-Britt Karlsson
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  2. Minna Pirhonen
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  3. Hannu T. Saarilahti
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  4. E. Tapio Palva
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Communicated by H. Hennecke

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Karlsson, MB., Pirhonen, M., Saarilahti, H.T. et al. Molecular cloning of ompRS, a regulatory locus controlling production of outer membrane proteins in Erwinia carotovora subsp. carotovora . Molec. Gen. Genet. 226, 353–360 (1991). https://doi.org/10.1007/BF00260646

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