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. 1978 Nov;136(2):538–547. doi: 10.1128/jb.136.2.538-547.1978

Repair of cross-linked DNA and survival of Escherichia coli treated with psoralen and light: effects of mutations influencing genetic recombination and DNA metabolism.

R R Sinden, R S Cole
PMCID: PMC218577  PMID: 361714

Abstract

Repair of cross-linked DNA was studied in Escherichia coli strains carrying mutations affecting DNA metabolism. In wild-type cells, DNA strands cut during cross-link removal were rejoined during a subsequent incubation into high-molecular-weight molecules. This rejoining was dependent on gene products involved in genetic recombination. A close correlation was found relating recombination proficiency, the rate of strand rejoining, and formation of viable progeny after DNA cross-linking by treatment with psoralen and light. Wild-type cells and other mutants which were Rec+ (sbcB, recL, recL sbcB, recB recC sbcA, recB recC sbcB, xthA1, and xthA11) rejoined cut DNA strands at a rate of 0.8 +/- 0.1 min -1 at 37 degrees C and survived 53 to 71 cross-links per chromosome. recB, recC, recB recC, recF, or polA strains showed reduced rates of strand rejoining and survived 4 to 13 cross-links per chromosome. Recombination-deficient strains (recA, recB recC sbcB recF, recB recL) and lexA failed to rejoin DNA strands after crosslink removal and were unable to form colonies after treatments producing as few as one to two cross-links per chromosome. Strand rejoining occurred normally in cells with mutations affecting DNA replication (dnaA, danB, dnaG, and dnaE) under both permissive and nonpermissive conditions for chromosome replication. In a polA polB dnaE strain strand rejoining occurred at 32 degree C but not at 42 degree C, indicating that some DNA synthesis was required for formation of intact recombinant molecules.

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Selected References

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  1. Barbour S. D., Nagaishi H., Templin A., Clark A. J. Biochemical and genetic studies of recombination proficiency in Escherichia coli. II. Rec+ revertants caused by indirect suppression of rec- mutations. Proc Natl Acad Sci U S A. 1970 Sep;67(1):128–135. doi: 10.1073/pnas.67.1.128. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Birge E. A., Low K. B. Detection of transcribable recombination products following conjugation in rec+, reCB- and recC-strains of Escherichia coli K12. J Mol Biol. 1974 Mar 15;83(4):447–457. doi: 10.1016/0022-2836(74)90506-3. [DOI] [PubMed] [Google Scholar]
  3. Broker T. R., Lehman I. R. Branched DNA molecules: intermediates in T4 recombination. J Mol Biol. 1971 Aug 28;60(1):131–149. doi: 10.1016/0022-2836(71)90453-0. [DOI] [PubMed] [Google Scholar]
  4. Buttin G., Wright M. Enzymatic DNA degradation in E. coli: its relationship to synthetic processes at the chromosome level. Cold Spring Harb Symp Quant Biol. 1968;33:259–269. doi: 10.1101/sqb.1968.033.01.030. [DOI] [PubMed] [Google Scholar]
  5. Capaldo-Kimball F., Barbour S. D. Involvement of recombination genes in growth and viability of Escherichia coli K-12. J Bacteriol. 1971 Apr;106(1):204–212. doi: 10.1128/jb.106.1.204-212.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Clark A. J. Progress toward a metabolic interpretation of genetic recombination of Escherichia coli and bacteriophage lambda. Genetics. 1974 Sep;78(1):259–271. doi: 10.1093/genetics/78.1.259. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Cole R. S. Inactivation of Escherichia coli, F' episomes at transfer, and bacteriophage lambda by psoralen plus 360-nm light: significance of deoxyribonucleic acid cross-links. J Bacteriol. 1971 Sep;107(3):846–852. doi: 10.1128/jb.107.3.846-852.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Cole R. S., Levitan D., Sinden R. R. Removal of psoralen interstrand cross-links from DNA of Escherichia coli: mechanism and genetic control. J Mol Biol. 1976 May 5;103(1):39–59. doi: 10.1016/0022-2836(76)90051-6. [DOI] [PubMed] [Google Scholar]
  9. Cole R. S. Light-induced cross-linking of DNA in the presence of a furocoumarin (psoralen). Studies with phage lambda, Escherichia coli, and mouse leukemia cells. Biochim Biophys Acta. 1970 Sep 17;217(1):30–39. doi: 10.1016/0005-2787(70)90119-x. [DOI] [PubMed] [Google Scholar]
  10. Cole R. S. Repair of DNA containing interstrand crosslinks in Escherichia coli: sequential excision and recombination. Proc Natl Acad Sci U S A. 1973 Apr;70(4):1064–1068. doi: 10.1073/pnas.70.4.1064. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Cole R. S., Sinden R. R. Repair of cross-linked DNA in Escherichia coli. Basic Life Sci. 1975;5B:487–495. doi: 10.1007/978-1-4684-2898-8_10. [DOI] [PubMed] [Google Scholar]
  12. Dall'Acqua F., Marciani S., Rodighiero G. Inter-strand cross-linkages occurring in the photoreaction between psoralen and DNA. FEBS Lett. 1970 Jul 29;9(2):121–123. doi: 10.1016/0014-5793(70)80330-1. [DOI] [PubMed] [Google Scholar]
  13. Emmerson P. T., West S. C. Identification of protein X of Escherichia coli as the recA+/tif+ gene product. Mol Gen Genet. 1977 Sep 21;155(1):77–85. doi: 10.1007/BF00268563. [DOI] [PubMed] [Google Scholar]
  14. Freifelder D. Molecular weights of coliphages and coliphage DNA. IV. Molecular weights of DNA from bacteriophages T4, T5 and T7 and the general problem of determination of M. J Mol Biol. 1970 Dec 28;54(3):567–577. doi: 10.1016/0022-2836(70)90127-0. [DOI] [PubMed] [Google Scholar]
  15. GEIDUSCHEK E. P. "Reversible" DNA. Proc Natl Acad Sci U S A. 1961 Jul 15;47:950–955. doi: 10.1073/pnas.47.7.950. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Ganesan A. K., Seawell P. C. The effect of lexA and recF mutations on post-replication repair and DNA synthesis in Escherichia coli K-12. Mol Gen Genet. 1975 Dec 1;141(3):189–205. doi: 10.1007/BF00341799. [DOI] [PubMed] [Google Scholar]
  17. Gefter M. L., Hirota Y., Kornberg T., Wechsler J. A., Barnoux C. Analysis of DNA polymerases II and 3 in mutants of Escherichia coli thermosensitive for DNA synthesis. Proc Natl Acad Sci U S A. 1971 Dec;68(12):3150–3153. doi: 10.1073/pnas.68.12.3150. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Holloman W. K., Radding C. M. Recombination promoted by superhelical DNA and the recA gene of Escherichia coli. Proc Natl Acad Sci U S A. 1976 Nov;73(11):3910–3914. doi: 10.1073/pnas.73.11.3910. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Horii Z., Clark A. J. Genetic analysis of the recF pathway to genetic recombination in Escherichia coli K12: isolation and characterization of mutants. J Mol Biol. 1973 Oct 25;80(2):327–344. doi: 10.1016/0022-2836(73)90176-9. [DOI] [PubMed] [Google Scholar]
  20. Hotchkiss R. D. Models of genetic recombination. Annu Rev Microbiol. 1974;28(0):445–468. doi: 10.1146/annurev.mi.28.100174.002305. [DOI] [PubMed] [Google Scholar]
  21. Howard-Flanders P., Theriot L. Mutants of Escherichia coli K-12 defective in DNA repair and in genetic recombination. Genetics. 1966 Jun;53(6):1137–1150. doi: 10.1093/genetics/53.6.1137. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. IYER V. N., SZYBALSKI W. A MOLECULAR MECHANISM OF MITOMYCIN ACTION: LINKING OF COMPLEMENTARY DNA STRANDS. Proc Natl Acad Sci U S A. 1963 Aug;50:355–362. doi: 10.1073/pnas.50.2.355. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Kanner L., Hanawalt P. Repair deficiency in a bacterial mutant defective in DNA polymerase. Biochem Biophys Res Commun. 1970 Apr 8;39(1):149–155. doi: 10.1016/0006-291x(70)90770-9. [DOI] [PubMed] [Google Scholar]
  24. Kirtikar D. M., Cathcart G. R., Goldthwait D. A. Endonuclease II, apurinic acid endonuclease, and exonuclease III. Proc Natl Acad Sci U S A. 1976 Dec;73(12):4324–4328. doi: 10.1073/pnas.73.12.4324. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Konrad E. B., Lehman I. R. Novel mutants of Escherichia coli that accumulate very small DNA replicative intermediates. Proc Natl Acad Sci U S A. 1975 Jun;72(6):2150–2154. doi: 10.1073/pnas.72.6.2150. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Kushner S. R., Nagaishi H., Clark A. J. Isolation of exonuclease VIII: the enzyme associated with sbcA indirect suppressor. Proc Natl Acad Sci U S A. 1974 Sep;71(9):3593–3597. doi: 10.1073/pnas.71.9.3593. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Kushner S. R., Nagaishi H., Templin A., Clark A. J. Genetic recombination in Escherichia coli: the role of exonuclease I. Proc Natl Acad Sci U S A. 1971 Apr;68(4):824–827. doi: 10.1073/pnas.68.4.824. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Lawley P. D., Brookes P. Molecular mechanism of the cytotoxic action of difunctional alkylating agents and of resistance to this action. Nature. 1965 May 1;206(983):480–483. doi: 10.1038/206480a0. [DOI] [PubMed] [Google Scholar]
  29. Lin P. F., Bardwell E., Howard-Flanders P. Initiation of genetic exchanges in lambda phage--prophage crosses. Proc Natl Acad Sci U S A. 1977 Jan;74(1):291–295. doi: 10.1073/pnas.74.1.291. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Marinus M. G., Adelberg E. A. Vegetative Replication and Transfer Replication of Deoxyribonucleic Acid in Temperature-Sensitive Mutants of Escherichia coli K-12. J Bacteriol. 1970 Dec;104(3):1266–1272. doi: 10.1128/jb.104.3.1266-1272.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Mount D. W., Low K. B., Edmiston S. J. Dominant mutations (lex) in Escherichia coli K-12 which affect radiation sensitivity and frequency of ultraviolet lght-induced mutations. J Bacteriol. 1972 Nov;112(2):886–893. doi: 10.1128/jb.112.2.886-893.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. REICH E., SHATKIN A. J., TATUM E. L. Bacteriocidal action of mitomycin C. Biochim Biophys Acta. 1961 Oct 14;53:132–149. doi: 10.1016/0006-3002(61)90800-9. [DOI] [PubMed] [Google Scholar]
  33. Radding C. M. Molecular mechanisms in genetic recombination. Annu Rev Genet. 1973;7:87–111. doi: 10.1146/annurev.ge.07.120173.000511. [DOI] [PubMed] [Google Scholar]
  34. Rothman R. H., Clark A. J. Defective excision and postreplication repair of UV-damaged DNA in a recL mutant strain of E. coli K-12. Mol Gen Genet. 1977 Oct 24;155(3):267–277. doi: 10.1007/BF00272805. [DOI] [PubMed] [Google Scholar]
  35. Rothman R. H., Clark A. J. The dependence of postreplication repair on uvrB in a recF mutant of Escherichia coli K-12. Mol Gen Genet. 1977 Oct 24;155(3):279–286. doi: 10.1007/BF00272806. [DOI] [PubMed] [Google Scholar]
  36. Rupp W. D., Howard-Flanders P. Discontinuities in the DNA synthesized in an excision-defective strain of Escherichia coli following ultraviolet irradiation. J Mol Biol. 1968 Jan 28;31(2):291–304. doi: 10.1016/0022-2836(68)90445-2. [DOI] [PubMed] [Google Scholar]
  37. Sedgwick S. G., Bridges B. A. Requirement for either DNA polymerase I or DNA polymerase 3 in post-replication repair in excision-proficient Escherichia coli. Nature. 1974 May 24;249(455):348–349. doi: 10.1038/249348a0. [DOI] [PubMed] [Google Scholar]
  38. Sinden R. R., Cole R. S. Topography and kinetics of genetic recombination in Escherichia coli treated with psoralen and light. Proc Natl Acad Sci U S A. 1978 May;75(5):2373–2377. doi: 10.1073/pnas.75.5.2373. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Vapnek D., Rupp W. D. Identification of individual sex-factor DNA strands and their replication during conjugation in thermosensitive DNA mutants of Escherichia coli. J Mol Biol. 1971 Sep 28;60(3):413–424. doi: 10.1016/0022-2836(71)90178-1. [DOI] [PubMed] [Google Scholar]
  40. Wechsler J. A., Gross J. D. Escherichia coli mutants temperature-sensitive for DNA synthesis. Mol Gen Genet. 1971;113(3):273–284. doi: 10.1007/BF00339547. [DOI] [PubMed] [Google Scholar]
  41. Wilkins B. M. Chromosome transfer from F-lac+ strains of Escherichia coli K-12 mutant at recA, recB, or recC. J Bacteriol. 1969 May;98(2):599–604. doi: 10.1128/jb.98.2.599-604.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Yajko D. M., Weiss B. Mutations simultaneously affecting endonuclease II and exonuclease III in Escherichia coli. Proc Natl Acad Sci U S A. 1975 Feb;72(2):688–692. doi: 10.1073/pnas.72.2.688. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Yoakum G. H., Cole R. S. Role of ATP in removal of psoralen cross-links from DNA of Escherichia coli permeabilized by treatment with toluene. J Biol Chem. 1977 Oct 25;252(20):7023–7030. [PubMed] [Google Scholar]
  44. Zieg J., Maples V. F., Kushner S. R. Recombinant levels of Escherichia coli K-12 mutants deficient in various replication, recombination, or repair genes. J Bacteriol. 1978 Jun;134(3):958–966. doi: 10.1128/jb.134.3.958-966.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]

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