Advertisement

Molecular and General Genetics MGG

, Volume 197, Issue 3, pp 461–466 | Cite as

Conjugative transfer of IncI1 plasmid DNA primase

  • Lee K. Chatfield
  • Brian M. Wilkins
Article

Summary

DNA primase of Collb-P9drd-1 generates RNA primers that are thought to initiate DNA synthesis on the conjugatively transferred strand of the plasmid. To examine whether plasmid-specified primase is transferred during conjugation, we exploited the property of the enzyme to promote bacterial DNA replication in dnaG (primase-defective) mutants of Escherichia coli. It was found that dnaG3 recipient cells, treated with rifampicin to inhibit transcription, recovered ability to synthesise bacterial DNA by a process requiring an active plasmid primase gene in donor cells and a functional conjugation system. A non-transferable primase gene in the donor strain complemented a primase-negative derivative of ColIb-P9drd-1, confirming that the enzyme responsible for recovery was supplied by donor cells. The implication is that certain proteins are transmitted from donor cells to promote conjugative metabolism of plasmid DNA in the recipients.

Keywords

Enzyme Rifampicin Donor Cell Recipient Cell Conjugation System 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Barth PT, Grinter NJ (1975) Assay of deoxyribonucleic acid homology using a single-strand-specific nuclease at 75 C. J Bacteriol 121:434–441Google Scholar
  2. Bayer ME (1979) The fusion sites between outer membrane and cytoplasmic membrane of bacteria: their role in membrane assembly and virus infection. In: Inouye M (ed) Bacterial outer membranes. Wiley-Interscience, New York, pp 167–202Google Scholar
  3. Boulnois GJ, Wilkins BM (1978) A ColI-specified product, synthesized in newly infected recipients, limits the amount of DNA transferred during conjugation of Escherichia coli K-12. J Bacteriol 133:1–9Google Scholar
  4. Boulnois GJ, Wilkins BM (1979) A novel priming system for conjugal synthesis of an IncIα plasmid in recipients. Mol Gen Genet 175:275–279Google Scholar
  5. Boulnois GJ, Beddoes MJ, Wilkins BM (1979) Rifampin disrupts conjugal and chromosomal deoxyribonucleic acid metabolism in Escherichia coli K-12 carrying some Inc1α plasmids. J Bacteriol 138:324–332Google Scholar
  6. Boulnois GJ, Wilkins BM, Lanka E (1982) Overlapping genes at the DNA primase locus of the large plasmid ColI. Nucl Acids Res 10:855–869Google Scholar
  7. Chatfield LK, Orr E, Boulnois GJ, Wilkins BM (1982) DNA primase of plasmid ColIb is involved in conjugal DNA synthesis in donor and recipient bacteria. J Bacteriol 152:1188–1195Google Scholar
  8. Clowes RC, Moody EEM (1966) Chromosomal transfer from “recombination-deficient” strains of Escherichia coli K-12. Genetics 53:717–726Google Scholar
  9. Davis EJ, Henry J (1982) Conjugal transfer replication of R64drd11 plasmid DNA in the donor cells of Escherichia coli K-12. Mol Gen Genet 187:305–309Google Scholar
  10. Falkow S, Tompkins LS, Silver RP, Guerry P, LeBlanc DJ (1971) The replication of R-factor DNA in Escherichia coli K-12 following conjugation. Ann NY Acad Sci 182:153–171Google Scholar
  11. Hillenbrand G, Morelli G, Lanka E, Scherzinger E (1978) Bacteriophage T7 DNA primase: a multifunctional enzyme involved in DNA replication. Cold Spring Harbor Symp Quant Biol 43:449–459Google Scholar
  12. Isaacson RE, Konisky J (1974) Studies on the regulation of colicin Ib synthesis I. Isolation of the ColIb-P9 plasmid. Mol Gen Genet 132:215–221Google Scholar
  13. Jeffreys AJ, Flavell RA (1977) A physical map of the DNA regions flanking the rabbit β-globin gene. Cell 12:429–439Google Scholar
  14. Kornberg A (1982) 1982 Supplement to DNA replication. W.H. Freeman and Co, San Francisco, pp S101-S125Google Scholar
  15. Lanka E, Fürste JP (1984) Function and properties of RP4 DNA primase. In: Hübscher U, Spadari S (eds) Proteins involved in DNA replication. Plenum Press, New York London, pp 265–280Google Scholar
  16. Lanka E, Scherzinger E, Günther E, Schuster H (1979) A DNA primase specified by I-like plasmids. Proc Natl Acad Sci USA 76:3632–3636Google Scholar
  17. Scherzinger E, Lanka E, Hillenbrand G (1977) Role of bacteriophage T7 DNA primase in the initiation of DNA strand synthesis. Nucl Acids Res 4:4151–4163Google Scholar
  18. Silver S, Ozeki H (1962) Transfer of deoxyribonucleic acid accompanying the transmission of colicinogenic properties by cell mating. Nature (London) 195:873–874Google Scholar
  19. Uemura H, Mizobuchi K (1982) Genetic and physical characterization of the ColIb plasmid using ColIb-R222 hybrids. Mol Gen Genet 185:1–12Google Scholar
  20. Vapnek D, Rupp WD (1970) Asymmetric segregation of the complementary sex-factor DNA strands during conjugation in Escherichia coli. J Mol Biol 53:287–303Google Scholar
  21. Welch MM, McHenry CS (1982) Cloning and identification of the product of the dnaE gene of Escherichia coli. J Bacteriol 152:351–356Google Scholar
  22. Wilkins BM (1975) Partial suppression of the phenotype of Escherichia coli K-12 dnaG mutants by some I-like conjugative plasmids. J Bacteriol 122:899–904Google Scholar
  23. Wilkins BM, Boulnois GJ, Lanka E (1981) A plasmid DNA primase active in discontinuous bacterial DNA replication. Nature (London) 290:217–221Google Scholar
  24. Wilkins BM, Hollom SE (1974) Conjugational synthesis of Flac + and ColI DNA in the presence of rifampicin and in Escherichia coli K12 mutants defective in DNA synthesis. Mol Gen Genet 134:143–156Google Scholar
  25. Willetts N, Wilkins B (1984) Processing of plasmid DNA during bacterial conjugation. Microbiol Rev 48:24–41Google Scholar

Copyright information

© Springer-Verlag 1984

Authors and Affiliations

  • Lee K. Chatfield
    • 1
  • Brian M. Wilkins
    • 1
  1. 1.Department of GeneticsUniversity of LeicesterLeicesterU.K.

Personalised recommendations