Skip to main content
SpringerLink
Log in

Investigations of the F conjugation gene traI:traI mutants and λtraI transducing phages

  • Published:
Molecular and General Genetics MGG Aims and scope Submit manuscript

Summary

A series of traI point and deletion mutants of Flac, and a traM mutant, were characterised. Complementation tests with an amber Flac traI mutant confirmed their genotypes, and in addition all the traI mutants, but not the traM mutant, were complemented by pRS31 (pSC101 traDI) and EDλ109 (λtraI). Judging from the efficiencies of plating of F-specific phages, none of the mutations affected pilus formation. The traI products of F and of the F-like plasmid R1 were interchangeable with each other but not with that of R100, while the traM product of F could not be replaced by those of R1 or of R100. Neither traI nor traM were needed for conjugal transfer of ColE1.

Three λ transducing phages carrying traI were isolated by in vivo or in vitro techniques, and characterised by genetic complementation tests, by analysis of the fragments produced by restriction endonucleases, and by measurement of heteroduplex molecules. The genetic structures together with the sizes and F coordinates, of the transfer regions carried by the phages were thereby determined.

Comparison of the proteins synthesised in UV-irradiated cells by one of the λraI phages with those made by a derivative carrying an amber traImutation, allowed the traI product to be identified as a protein of molecular weight 174,000. In addition, the molecular weights of the traD (84,000), traS (18,000), and traT (25,000) products made by the λtraSTDI phage EDλ107 were measured.

The possible roles of the traI and traM products in conjugation are discussed.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Achtman, M., Skurray, R.: A redefinition of the mating phenomenon in bacteria. In: Microbial interactions. Vol. 3, pp. 232–279 (J.L. Reissig, ed.) London: Pub. Chapman and Hall 1977

    Google Scholar 

  • Achtman, M., Skurray, R.A., Thompson, R., Helmuth, R., Hall, S., Beutin, L., Clark, A.J.: Assignment of tra cistrons to EcoRI fragments of F sex factor DNA. J. Bacteriol. 133, 1383–1392 (1978)

    Google Scholar 

  • Achtman, M., Willetts, N.S., Clark, A.J.: Beginning a genetic analysis of conjugational transfer determined by the F factor in E.coli by isolation and characterisation of transfer-deficient mutants. J. Bacteriol. 106, 529–538 (1971)

    Google Scholar 

  • Achtman, M., Willetts, N.S., Clark, A.J.: A conjugational complementation analysis of transfer-deficient mutants of Flac in E.coli. J. Bacteriol. 110, 831–842 (1972)

    Google Scholar 

  • Alfaro, G., Willetts, N.S.: The relationship between the transfer systems of some bacterial plasmids. Genet. Res. 20, 279–289 (1972)

    Google Scholar 

  • Blattner, F.R., Fiandt, M., Hass, K.K., Twose, P.A., Szybalski, W.: Deletions and insertions in the immunity region of coliphage lambda: revised measurement of the promoter-startpoint distance. Virology 62, 458–471 (1974)

    Google Scholar 

  • Childs, G.J., Ohtsubo, H., Ohtsubo, E., Sonnenberg, F., Freundlich, M.: Restriction endonuclease mapping of the E.coli K12 chromosome in the vicinity of the ilv genes. J. Mol. Biol. 117, 175–193 (1977)

    Google Scholar 

  • Dempsey, W.B., Willetts, N.S.: Plasmid cointegrates of prophage lambda and R factor R100. J. Bacteriol. 126, 166–176 (1976)

    Google Scholar 

  • Egawa, R., Hirota, Y.: Inhibition of fertility by multiple drug-resistance factor in E.coli K12. Japan.J. Genet. 37, 66–69 (1962)

    Google Scholar 

  • Finnegan, D.J., Willetts, N.S.: Two classes of Flac mutants insensitive to transfer inhibition by an F-like R factor. Mol. Gen. Genet. 111, 256–264 (1971)

    Google Scholar 

  • Finnegan, D.J., Willetts, N.S.: The nature of the transfer inhibitor of several F-like plasmids. Mol. Gen. Genet. 119, 57–66 (1972)

    Google Scholar 

  • Finnegan, D.J., Willetts, N.S.: The site of action of the F transfer inhibitor. Mol. Gen. Genet. 127, 307–316 (1973)

    Google Scholar 

  • Foster, T.J., Willetts, N.S.: Genetic analysis of deletions of R100-1 that are both transfer-deficient and tetracycline-sensitive. J. Gen. Microbiol. 93, 133–140 (1976)

    Google Scholar 

  • Foster, T.J., Willetts, N.S.: Characterisation of transfer-deficient mutants of the R100-1 TcS plasmid pDU202, caused by insertion of Tn10. Mol. Gen. Genet. 156, 107–114 (1977)

    Google Scholar 

  • Gasson, M.J., Willetts, N.S.: Further characterisation of the F fertility inhibition systems of “unusual” Fin+ plasmids. J. bacteriol. 131, 413–420 (1977)

    Google Scholar 

  • Georgopoulos, C.P., Hendrix, R.W., Casjens, S.R., Kaiser, A.D.: Host participation in bacteriophage lambda head assembly. J. Mol. Biol. 76, 45–60 (1973)

    Google Scholar 

  • Haggerty, D.M., Schleif, R.F.: Location in bacteriophage lambda DNA of cleavage sites of the site-specific endonuclease from B.amyloliquefaciens H. J. Virol. 18, 659–663 (1976)

    Google Scholar 

  • Hashimoto, H., Hirota, Y.: Gene recombination and segregation of resistance factor R in E.coli. J. Bacteriol. 91, 51–62 (1966)

    Google Scholar 

  • Helmuth, R., Achtman, M.: Operon structure of DNA transfer cistrons on the F sex factor. Nature 257, 652–656 (1975)

    Google Scholar 

  • Hendrix, R.W.: Identification of proteins coded in phage lambda. In: The bacteriophage lambda (A.D. Hershey, ed.), pp. 355–370 New York: Cold Spring Harbor Laboratory 1971

    Google Scholar 

  • Hu, S., Ptashne, K., Cohen, S.N., Davidson, N.: αβ sequence of F is IS3. J. Bacteriol. 123, 687–692 (1975)

    Google Scholar 

  • Ippen-Ihler, K., Achtman, M., Willetts, N.S.: A deletion map of the E.coli K12 sex factor F: the order of eleven transfer cistrons. J. Bacteriol. 110, 857–863 (1972)

    Google Scholar 

  • Jaskunas, S.R., Lindahl, L., Nomura, M., Burgess, R.R.: Identification of two copies of the gene for the elongation factor EF-Tu in E.coli. Nature 257, 458–462 (1975)

    Google Scholar 

  • Kamp, D., Kahmann, R., Zipser, D., Roberts, R.J.: Mapping of restriction sites in the attachment site region of bacteriophage lambda. Mol. Gen. Genet. 154, 231–248 (1977)

    Google Scholar 

  • Kennedy, M., Beutin, L., Achtman, M., Skurray, R., Rahmsdorf, U., Herrlich, P.: Conjugation proteins encoded by the F sex factor. Nature 270, 580–585 (1977)

    Google Scholar 

  • Kingsman, A. Willetts, N.: The requirements for conjugal DNA synthesis in the donor strain during Flac transfer. J. Mol. Biol. 122, 287–300 (1978)

    Google Scholar 

  • Kirschbaum, J.B., Scaife, J.G.: Evidence for a λ transducing phage carrying the genes for the β and β′ subunits of E.coli RNA polymcrase. Mol. Gen. Genet. 132, 193–201 (1974)

    Google Scholar 

  • Krell, K., Gottesman, M.E., Parks, J.S., Eisenberg, M.A.: Escape synthesis of the biotin operon in induced λb2 lysogens. J. Mol. Biol. 68, 69–82 (1972)

    Google Scholar 

  • Laemmli, U.K.: Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227, 680–685 (1970)

    PubMed  Google Scholar 

  • Landy, A., Ross, W.: Viral integration and excision: structure of the lambda att sites. Science 197, 1147–1160 (1977)

    Google Scholar 

  • Lederberg, E.M., Cohen, S.N.: Transformation of S.typhimurium by plasmid DNA. J. Bacteriol. 119, 1072–1074 (1974)

    Google Scholar 

  • McIntire, S., Willetts, N.S.: Plasmid cointegrates of Flac and lambda prophage. J. Bacteriol. 134, 184–192 (1978a)

    Google Scholar 

  • McIntire, S., Willetts, N.S.: Transfer-deficient cointegrates of Flac and lambda prophage. J. Bacteriol. (1978b) submitted for publication.

  • Meynell, E., Datta, N.: Mutant drug resistance factors of high transmissibility. Nature 214, 885–887(1967)

    Google Scholar 

  • Miki, T., Horiuchi, T., Willetts, N.S.: Identification and characterisation of four new tra cistrons on the E.coli sex factor F. Plasmid 1, 316–323 (1978)

    Google Scholar 

  • Ohtusubo, E., Nishimura, Y., Hirota, Y.: Transfer-defective mutants of sex factors in E.coli. I. Defective mutants and complementation analysis. Genetics 64, 173–188 (1970)

    Google Scholar 

  • Sharp, P.A., Cohen, S.N., Davidson, N.: Electron microscope heteroduplex studies of sequence relations among plasmids of E.coli. II. Structure of drug resistance (R) factors and F factors. J. Mol. Biol. 75, 235–255 (1973)

    Google Scholar 

  • Sharp, P.A., Hsu, M., Ohtsubo, E., Davidson, N.: Electron microscope heteroduplex studies of sequence relations among plasmids of E.coli. I. Structure of F prime factors. J. Mol. Biol. 71, 471–497 (1972)

    Google Scholar 

  • Skurray, R.A., Nagaishi, H., Clark, A.J.: Molecular cloning of DNA from F sex factor of Escherichia coli K12. Proc. Natl. Acad. Sci. U.S.A. 73, 64–68 (1976)

    Google Scholar 

  • Studier, F.W.: Analysis of bacteriophage T7 early RNAs and proteins on slab gels. J. Mol. Biol. 79, 237–248 (1973)

    Google Scholar 

  • Warren, G., Sherratt, D.: Complementation of transfer deficient ColEl mutants. Mol. Gen. Genet. 151, 197–201 (1977)

    Google Scholar 

  • Willetts, N.S.: The plasmid-specificity of two proteins required for conjugation in E.coli K12. Nature New Biol. 230, 183–185 (1971)

    Google Scholar 

  • Willetts, N.S.: Recombination and the E.coli K12 sex factor F. J. Bacteriol. 121, 36–43 (1975)

    Google Scholar 

  • Willetts, N.S.: The transcriptional control of fertility in F-like plasmids. J. Mol. Biol. 112, 141–148 (1977a)

    Google Scholar 

  • Willetts, N.S.: Genetics of conjugation. In: R factor-drug resistance plasmid (S. Mitsuhashi, ed.), pp. 89–107. Tokyo: University of Tokyo Press 1977

    Google Scholar 

  • Willetts, N.S.: Control of conjugation in F-like plasmids. In: Microbiology (D. Schlessinger, ed.), pp. 211–213. A.S.M., Washington D.C. 1978

    Google Scholar 

  • Willetts, N.S., Achtman, M.: A genetic analysis of transfer by the E.coli sex factor F, using Pl transductional complementation. J. Bacteriol. 110, 843–851 (1972)

    Google Scholar 

  • Willetts, N.S., Finnegan, D.J.: Characteristics of E.coli K12 strains carrying both an F prime and an R factor. Genet. Res. 16, 113–122 (1970)

    Google Scholar 

  • Willetts, N.S., McIntire, S.: The exploitation of λtra transducing phages in the study of bacterial conjugation. In: Contributions to microbiology and immunology (23rd OHOLO Conference). Basel: Karger A.G. 1978a (in press)

    Google Scholar 

  • Willetts, N.S., McIntire, S.: Isolation and characterisation of λtra transducing phages from EDFL223 (Flac traB::EDλ4). J. Mol. Biol. (1978b) (in press)

  • Willetts, N.S., Moore, P.M., Paranchych, W.: Variant pili produced by Flac mutants and derepressed F-like plasmids. J. Bacteriol. (1979) submitted for publication

Download references

Author information

Authors and Affiliations

  1. Department of Molecular Biology, University of Edinburgh, EH9 3JR, Edinburgh, Scotland

    Neil Willetts & John Maule

Authors
  1. Neil Willetts
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. John Maule
    View author publications

    You can also search for this author in PubMed Google Scholar

Additional information

Communicated by W. Arber

Rights and permissions

Reprints and permissions

About this article

Cite this article

Willetts, N., Maule, J. Investigations of the F conjugation gene traI:traI mutants and λtraI transducing phages. Molec. Gen. Genet. 169, 325–336 (1979). https://doi.org/10.1007/BF00382278

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00382278

Keywords

Search

Navigation