Skip to main content
Log in

The DNa-protein relaxation complex of the plasmid RK2: Location of the site-specific nick in the region of the proposed origin of transfer

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

Summary

The broad host range plasmid, RK2, has been isolated as a DNA-protein relaxation complex. Nicking of the plasmid DNA in the relaxation complex occurs at a single specific site (rlx) located approximately 20 kb away from the origin of DNA replication. A cis-acting function required for plasmid transfer, the presumptive origin of transfer, maps in the same region as rlx. The region of RK2 encompassing rlx has been cloned onto pBR322 and shown to promote mobilization of the hybrid plasmid by an RK2 derivative. These results indicate that the RK2 relaxation complex nicks at or near the origin of transfer of the RK2 plasmid.

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

Access this article

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

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Barth, P.T., Grinter, N.J., Bradley, D.E.: Conjugal transfer system of plasmid RP4: analysis by transposon 7 insertion. J. Bacteriol. 133, 43–52 (1978)

    Google Scholar 

  • Blair, D., Helinski, D.R.: Relaxation complexes of plasmid DNA and protein. I. Strand-specific association of protein and DNA in the relaxed complexes of plasmids ColE1 and ColE2. J. Biol. Chem. 250, 8785–8789 (1975)

    Google Scholar 

  • Bolivar, F., Rodriguez, R.L., Greene, P.Y., Betlach, M.C., Heynecker, H.L., Boyer, H.W., Crosa, Y.H., Falkow, S.: Construction and characterization of new cloning vehicles. II. A multiple cloning system. Gene 2, 95–113 (1977)

    Google Scholar 

  • Clewell, D., Helinski, D.R.: Supercoiled circular DNA-protein complex in Escherichia coli: purification and induced conversion to an open circular DNA form. Proc. Natl. Acad. Sci. U.S.A. 62, 1159–1166 (1969)

    Google Scholar 

  • Crisona, N.J., Clark, A.J.: Increase in conjugational transmission frequency of nonconjugative plasmids. Science 196, 186–187 (1977)

    Google Scholar 

  • Datta, N., Hedges, R.W.: Host ranges of R factors. J. Gen. Microbiol. 70, 453–460 (1972)

    Google Scholar 

  • Dougan, G., Sherratt, D.: The transposon Tnl as a probe for studying ColEl structure and function. Mol. Gen. Genet. 151, 151–160 (1977)

    Google Scholar 

  • Figurski, D., Meyer, R., Miller, D.S., Helinski, D.R.: Generation in vitro of deletions in the broad host range plasmid RK2 using phage Mu insertions and a restriction endonuclease. Gene 1, 107–119 (1976)

    Google Scholar 

  • Guiney, D.G., Helinski, D.R.: Relaxation complexes of plasmid DNA and protein. III. Association of protein with the 5′ terminus of the broken DNA strand in the relaxed complex of plasmid ColE1. J. Biol. Chem. 250, 8796–8803 (1975)

    Google Scholar 

  • Holloway, B.W.: Plasmids that mobilize bacterial chromosome. Plasmid 2, 1–19 (1979)

    Google Scholar 

  • Ingram, L.C., Richmond, M.H., Sykes, R.B.: Molecular characterization of the R factors implicated in the carbenicillin resistance of a sequence of Pseudomonas aeruginosa strains isolated from burn. Antimicrob. Agents, Chemother. 3, 279–288 (1973)

    Google Scholar 

  • Inselberg, J.: Studies of colicin E1 plasmid functions by analysis of deletions and TnA insertions of the plasmid. J. Bacteriol. 132, 332–340 (1977)

    Google Scholar 

  • Kline, B.C., Helinski, D.R.: F1 sex factor of Escherichia coli. Size and purification in the form of a strand-specific relaxation complex of supercoiled deoxyribonucleic acid and protein. Biochemistry 10, 4975–4980 (1971)

    Google Scholar 

  • Kuperstoch-Portnoy, Y.M., Miklos, G.L., Helinski, D.R.: Properties of the relaxation complexes of supercoiled deoxyribonucleic acid and protein of the R plasmids R64, R28K, and R6K. J. Bacteriol. 120, 545–548 (1974)

    Google Scholar 

  • Lovett, M.A., Guiney, D.G., Helinski, D.R.: Relaxation complexes of plasmids ColE1 and ColE2: Unique site of the nick in the open circular DNA of the relaxed complexes. Proc. Natl. Acad. Sci. USA, 71, 3854–3857 (1974)

    Google Scholar 

  • Lovett, M.A., Helinski, D.R.: Relaxation complexes of plasmid DNA and protein. II. Characterization of the proteins associated with the unrelaxed and relaxed complexes of plasmid ColE1. J. Biol. Chem. 250, 8790–8795 (1975)

    Google Scholar 

  • Lovett, M.A., Sparks, R.B., Helinski, D.R.: Bidirectional replication of plasmid R6K DNA in Escherichia coli; correspondence between origin of replication and position of single-strand break in relaxed complex. Proc. Natl. Acad. Sci. U.S.A. 72, 2905–2909 (1975)

    Google Scholar 

  • Meyer, R., Figurski, D., Helinski, D.R.: Physical and genetic studies with restriction endonucleases on the broad host-range plasmid RK2. Mol. Gen. Genet. 152, 129–135 (1977)

    Google Scholar 

  • Meyer, R.J., Helinski, D.R.: Unidirectional replication of the P-group plasmid RK2. Biochim. Biophys. Acta 478, 109–113 (1977)

    Google Scholar 

  • Morris, C.F., Hershberger, C.L., Rownd, R.: Strand-specific nick in open circular R-factor deoxyribonucleic acid: Attachment of the linear strand to a proteinaceous cellular component. J. Bacteriol. 114, 300–308 (1973)

    Google Scholar 

  • Olsen, R.H., Shipley, P.: Host ranges and properties of the Pseudomonas aeruginosa R factor R1822. J. Bacteriol. 113, 772–780 (1973)

    Google Scholar 

  • Studier, F.W.: Sedimentation studies of the size and shape of DNA. J. Mol. Biol 11, 373–390 (1965)

    Google Scholar 

  • Thompson, R., Achtman, M.: The control region of the F sex factor DNA transfer cistrons: Restriction mapping and DNA cloning. Mol. Gen. Genet. 165, 295–304 (1978)

    Google Scholar 

  • Tomizawa, J., Ohmori, H., Bird, R.: Origin of replication of colicin E1 plasmid DNA. Proc. Natl. Acad. Sci. U.S.A. 74, 1865–1869 (1977)

    Google Scholar 

  • Warren, G.J., Twigg, A.J., Sherratt, D.J.: ColE1 plasmid mobility and relaxation complex. Nature 274, 259–261 (1978)

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Additional information

Communicated by E. Bautz

Rights and permissions

Reprints and permissions

About this article

Cite this article

Guiney, D.G., Helinski, D.R. The DNa-protein relaxation complex of the plasmid RK2: Location of the site-specific nick in the region of the proposed origin of transfer. Molec. Gen. Genet. 176, 183–189 (1979). https://doi.org/10.1007/BF00273212

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00273212

Keywords

Navigation