Molecular and General Genetics MGG

, Volume 195, Issue 1–2, pp 267–274 | Cite as

Repair of DNA double-strand breaks in Escherichia coli K12 requires a functional recN product

  • Steven M. Picksley
  • Paul V. Attfield
  • Robert G. Lloyd


Mutation of the recN gene of Escherichia coli in a recBC sbcB genetic background blocks conjugational recombination and confers increased sensitivity to UV light and mitomycin C. The basis for this phenotype was investigated by monitoring the properties associated with recN mutations in otherwise wild-type strains. It was established that recN single mutants are almost fully resistant to UV irradiation, and that there is no detectable defect in repair of UV lesions by excision, error-prone, or recombinational mechanisms. However, recN mutations confer sensitivity to mitomycin C and ionizing radiation both in wild-type and recB sbcB strains. The sensitivity to ionizing radiation is correlated with a deficiency in the capacity to repair DNA double-strand breaks by a UV inducible mechanism. Recombinant λ phages that complement the recombination and repair defects of recN recBC sbcB mutants have been identified, and the recN gene has been cloned from these phages into a low copy-number plasmid.


Escherichia Coli Recombination Detectable Defect Single Mutant Repair Defect 
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.


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  1. Arthur HM, Bramhill D, Eastlake P, Emmerson PT (1982) Cloning of the uvrD gene of E. coli and identification of the product. Gene 19:285–295Google Scholar
  2. Bachmann BJ (1972) Pedigrees of some mutant strains of Escherichia coli K-12. Bacteriol Rev 36:525–557Google Scholar
  3. Bachmann BJ (1983) Linkage map of Escherichia coli K12, edition 7. Microbiol Rev 47:180–230Google Scholar
  4. Berman ML, Enquist LW, Silhavy TJ (1981) Advanced bacterial genetics. Cold Spring Harbor Laboratory, New YorkGoogle Scholar
  5. Birge EA, Low KB (1974) Detection of transcribable recombination products following conjugation in Rec+, RecB-, and RecC- strains of Escherichia coli K12. J Mol Biol 83:447–457Google Scholar
  6. Bonura T, Smith KC (1975) Enzymic production of deoxyribonucleic acid double-strand breaks after ultraviolet irradiation of Escherichia coli K-12. J Bacteriol 121:511–517Google Scholar
  7. Clark AJ (1973) Recombination deficient mutants of E. coli and other bacteria. Ann Rev Genet 7:67–86Google Scholar
  8. Clark AJ, Margulies AD (1965) Isolation and characterisation of recombination — deficient mutants of Escherichia coli K12. Proc Natl Acad Sci USA 53:451–459Google Scholar
  9. DasGupta C, Radding CM (1982) Polar branch migration promoted by recA protein: Effect of mismatched base pairs. Proc Natl Acad Sci USA 79:762–766Google Scholar
  10. Ganesan AK (1974) Persistence of pyrimidine dimers during postreplication repair of ultraviolet light-irradiated Escherichia coli J Molec Biol 87:103–119Google Scholar
  11. Guyer MS (1978) The γδ sequence of F is an insertion sequence. J Mol Biol 126:347–365Google Scholar
  12. Hall JD, Mount DW (1981) Mechanisms of DNA replication and mutagenesis in ultraviolet-irradiated bacteria and mammalian cells. Prog Nucleic Acid Res Mol Biol 25:54–126Google Scholar
  13. Hanawalt PC, Cooper PK, Ganesan AK, Smith CA (1979) DNA repair in bacterial and mammalian cells. Ann Rev Biochem 48:783–836Google Scholar
  14. Horii ZI, Clark AJ (1973) Genetic analysis of the RecF pathway to genetic recombination in Escherichia coli K12: isolation and characterisation of mutants. J Mol Biol 80:327–344Google Scholar
  15. Ish-Horowicz D, Burke JF (1981) Rapid and efficient cosmid cloning. Nucleic Acids Res 9:2989–2998Google Scholar
  16. Krasin F, Hutchinson F (1977) Repair of DNA double-strand breaks in Escherichia coli which requires recA function and the presence of a duplicate genome. J Mol Biol 116:81–98Google Scholar
  17. Krasin F, Hutchinson F (1981) Repair of DNA double strand breaks in Escherichia coli cells requires synthesis of proteins that can be induced by UV light. Proc Natl Acad Sci USA 78:3450–3453Google Scholar
  18. Kushner SR, Nagaishi H, Tomplin A, Clark AJ (1971) Genetic recombination in Escherichia coli: The role of Exonuclease I. Proc Natl Acad Sci USA 68:824–827Google Scholar
  19. Kushner SR, Nagaishi H, Clark AJ (1972) Indirect suppression of recB and recC mutations by Exonuclease I deficiency. Proc Natl Acad Sci USA 69:1366–1370Google Scholar
  20. Lin PF, Bardwell E, Howard-Flanders P (1977) Initiation of genetic exchanges in phage-prophage crosses. Proc Natl Acad Sci USA 74:291–295Google Scholar
  21. Little JW, Mount DW (1982) The SOS regulatory system of Escherichia coli. Cell 29:11–22Google Scholar
  22. Livneh Z, Lehman IR (1982) Recombinational bypass of pyrimidine dimers promoted by the recA protein of Escherichia coli. Proc Natl Acad Sci USA 79:3171–3175Google Scholar
  23. Lloyd RG, Barbour SD (1974) The genetic location of the sbcA gene of Escherichia coli. Mol Gen Genet 134:157–171Google Scholar
  24. Lloyd RG, Low B (1976) Some genetic consequence of changes in the level of recA gene function in Escherichia coli K12. Genetics 84:675–694Google Scholar
  25. Lloyd RG, Low B, Godson GN, Birge EA (1974) Isolation and characterisation of an Escherichia coli K12 mutant with a temperature-sensitive RecA- phenotype. J Bacteriol 120:407–415Google Scholar
  26. Lloyd RG, Picksley SM, Prescott C (1983) Inducible expression of a gene specific to the RecF pathway for recombination in Escherichia coli K12. Mol Gen Gent 190:162–167Google Scholar
  27. Lloyd RG, Benson FE, Shurvinton CE (1984) Effect of ruv mutations on recombination and DNA repair in Escherichia coli. Mol Gen Genet 194:303–309Google Scholar
  28. Miller JH (1972) Experiments in molecular genetics. Cold Spring Harbour Laboratory, New YorkGoogle Scholar
  29. Pollard EC, Fluke DJ, Kazanis D (1981) Induced radioresistance: an aspect of induced repair. Mol Gen Genet 184:421–429Google Scholar
  30. Prakash S, Prakash L, Burke W, Montelone BA (1980) Effects of the RAD52 gene on recombination in Saccharomyces cerevisiae. Genetics 94:31–50Google Scholar
  31. Radding C (1982) Homologous pairing and strand exchange in genetic recombination. Ann Rev Genet 16:405–437Google Scholar
  32. Ream LW, Margossian L, Clark AJ, Hansen FG, von Meyenburg K (1980) Genetic and physical mapping of recF in Escherichia coli K-12. Mol Gen Genet 180:115–121Google Scholar
  33. Resnick MA, Martin P (1976) The repair of double strand breaks in the nuclear DNA of Saccharomyces cerevisiae and its genetic control. Mol Gen Genet 143:119–129Google Scholar
  34. Rothman RH, Clark AJ (1977) The dependence of postreplication repair on uvrB in a recF mutant of Escherichia coli K12. Mol Gen Genet 155:279–286Google Scholar
  35. Rupp WD, Wilde CE, Reno DL, Howard-Flanders P (1971) Echanges between DNA strands in ultraviolet-irradiated Escherichia coli. J Mol Biol 61:25–44Google Scholar
  36. Sancar A, Rupp WD (1983) A novel repair enzyme: UVRABC excision nuclease of Escherichia coli cuts a DNA strand on both sides of the damaged region. Cell 33:249–260Google Scholar
  37. Shurvinton CE, Lloyd RG (1982) Damage to DNA induces expression of the ruv gene of Escherichia coli. Mol Gen Genet 185:352–355Google Scholar
  38. Shurvinton CE, Lloyd RG, Benson FE, Attfield PV (1984) Genetic analysis and molecular cloning of the Escherichia coli ruv gene. Mol Gen Genet 194:322–329Google Scholar
  39. Sinden RR, Cole RS (1978) Topography and kinetics of genetic recombination in Escherichia coli treated with psoralen and light. Proc Natl Acad Sci USA 75:2373–2377Google Scholar
  40. Szostak JW, Orr-Weaver TL, Rothstein RJ, Stahl FW (1983) The double-strand break repair model for recombination. Cell 33:25–35Google Scholar
  41. Timmis KN (1981) Gene manipulation in vitro. In Glover SW, Hopwood DA (eds), Genetics as a tool in microbiology, Cambridge University Press pp 49–109Google Scholar
  42. Wang TV, Smith KC (1982) Effects of the ssb-1 and ssb-113 mutations on survival and DNA repair in UV-irradiated uvrB strains of Escherichia coli K-12Google Scholar
  43. West SC, Cassuto E, Howard-Flanders P (1982) Post-replication repair in E. coli: Strand exchange reactions of gapped DNA by RecA protein. Mol Gen Genet 187:209–217Google Scholar
  44. Willetts NS, Mount DW (1969) Genetic analysis of rec - mutants of Escherichia coli carrying rec mutations cotransducible with thyA. J Bacteriol 100:923–934Google Scholar
  45. Witkin EM (1976) Ultraviolet mutagenesis and inducible deoxyribonucleic acid repair in Escherichia coli. Bacteriol Rev 40:869–907Google Scholar

Copyright information

© Springer-Verlag 1984

Authors and Affiliations

  • Steven M. Picksley
    • 1
  • Paul V. Attfield
    • 1
  • Robert G. Lloyd
    • 1
  1. 1.Genetics DepartmentUniversity of NottinghamNottinghamEngland

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