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Mutants that overproduce TraTp, a plasmid-specified major outer membrane protein of Escherichia coli

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Summary

The isolation of a series of plasmid mutant derivatives that overproduce the traT outer membrane protein, TraTp, is described. Some of the mutants directed the synthesis of 10-fold more TraTp (200,000 copies/cell) than did the parental plasmid (20,000 copies/cell). The proteins specified by all mutant plasmids except one were correctly inserted into the outer membrane and exposed on the cell surface. The TraTp that was not correctly inserted did not mediate the expected levels of surface exclusion and serum resistance, suggesting that surface localization is a requirement of TraTp function. The overproduction of TraTp was deleterious to bacterial growth, particularly that of minicell mutants of E. coli K-12.

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References

  • Achtman M, Kusecek B, Timmis KN (1978) tra cistrons and proteins encoded by the Escherichia coli antibiotic resistance plasmid R6-5. Mol Gen Genet 163:169–179

    Google Scholar 

  • Achtman M, Manning PA, Kusecek B, Schwuchow S, Willetts N (1980) A genetic analysis of F sex factor cistrons needed for surface exclusion in Escherichia coli. J Mol Biol 138:779–795

    Google Scholar 

  • Achtman M, Willetts NS, Clark AJ (1971) Beginning a genetic analysis of conjugational transfer determined by the F factor of Escherichia coli by isolation and characterization of transfer deficient mutants. J Bacteriol 106:529–538

    Google Scholar 

  • Andres I, Slocombe PM, Cabello F, Timmis JK, Lurz R, Burkardt HJ, Timmis KN (1979) Plasmid replication functions. II. Cloning analysis of the repA replication region of antibiotic resistance plasmid R6-5. Mol Gen Genet 168:1–25

    Google Scholar 

  • Chang ACY, Cohen SN (1978) Construction and characterization of amplifiable multicopy DNA cloning vehicles derived from the p15a cryptic miniplasmid. J Bacteriol 134:1141–1156

    Google Scholar 

  • Cohen SN, Chang ACY, Hsu L (1972) Non-chromosomal antibiotic resistance in bacteria: genetic transformation of Escherichia coli by R factor DNA. Proc Natl Acad Sci USA 69:2110–2114

    Google Scholar 

  • Ferrazza D, Levy SB (1981) Biochemical and immunological characterization of an R plasmid-encoded protein with properties resembling those of major cellular outer membrane proteins. J Bacteriol 144:149–148

    Google Scholar 

  • Halegoua S, Inouye M (1979) Biosynthesis and assembly of the outer membrane proteins. In: Inouye M (ed) Bacterial Outer Membranes: Biogenesis and Functions. New York, Wiley and Sons, pp 67–113

    Google Scholar 

  • Henning U, Royer H-D, Teather RM, Hindennach I, Hollenberg CP (1979) Cloning of the structural gene (ompA) for an integral outer membrane protein of Escherichia coli K-12. Proc Natl Acad Sci USA 76:4360–4364

    Google Scholar 

  • Hoar DK (1970) Fertility regulation of F like resistance transfer factors. J Bacteriol 101:916–920

    Google Scholar 

  • Humphreys GO, Willshaw CA, Smith HR, Anderson ES (1976) Mutagenesis of plasmid DNA with hydroxylamine: isolation of mutants of multicopy plasmids. Mol Gen Genet 145:101–108

    Google Scholar 

  • Manning PA, Beutin L, Achtman M (1980) Outer membrane of Escherichia coli: properties of the F sex factor traT protein which is involved in surface exclusion. J Bacteriol 142:285–294

    Google Scholar 

  • Moll A, Cabello F, Timmis KN (1979) Rapid assay for the determination of bacterial resistance to the lethal activity of serum. FEMS Microbiol Lett 6:273–276

    Google Scholar 

  • Moll A, Manning PA, Timmis KN (1980) Plasmid-determined resistance to serum bactericidal activity: a major outer membrane protein, the traT gene product, is responsible for plasmid-specified serum resistance in Escherichia coli. Infect Immun 28:359–367

    Google Scholar 

  • Ogata RT, Levine RP (1980) Characterization of complement resistance in Escherichia coli conferred by the antibiotic resistance plasmid R100. J Immunol 125:1494–1498

    Google Scholar 

  • Osborn MJ, Wu HCP (1980) Proteins of the outer membrane of Gram negative bacteria. Annu Rev Microbiol 34:369–422

    Google Scholar 

  • Timmis KN, Cabello F, Cohen SN (1978) Cloning and characterization of EcoRI and HindIII restriction endonuclease-generated fragments of antibiotic resistance plasmids R6-5 and R6. Mol Gen Genet 162:121–137

    Google Scholar 

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Communicated by J. Schell

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Manning, P.A., Timmis, J.K., Moll, A. et al. Mutants that overproduce TraTp, a plasmid-specified major outer membrane protein of Escherichia coli . Molec Gen Genet 187, 426–431 (1982). https://doi.org/10.1007/BF00332623

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  • DOI: https://doi.org/10.1007/BF00332623

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