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.
Similar content being viewed by others
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
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
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
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
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
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
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
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
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
Hoar DK (1970) Fertility regulation of F like resistance transfer factors. J Bacteriol 101:916–920
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
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
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
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
Ogata RT, Levine RP (1980) Characterization of complement resistance in Escherichia coli conferred by the antibiotic resistance plasmid R100. J Immunol 125:1494–1498
Osborn MJ, Wu HCP (1980) Proteins of the outer membrane of Gram negative bacteria. Annu Rev Microbiol 34:369–422
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
Author information
Authors and Affiliations
Additional information
Communicated by J. Schell
Rights and permissions
About this article
Cite this article
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
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00332623