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. 1972 Dec;112(3):1237–1246. doi: 10.1128/jb.112.3.1237-1246.1972

Excision Repair Characteristics of recBres and uvrC Strains of Escherichia coli

Takesi Kato a,1
PMCID: PMC251554  PMID: 4344920

Abstract

An Escherichia coli strain carrying the recB21 and res-1 mutations showed an abnormally low level of colony-forming ability although it grew essentially normally in liquid medium. The recB21 res-1 strain showed little, if any, of the ultraviolet (UV)-induced deoxyribonucleic acid (DNA) breakdown characteristic of the res-1 mutant. Nevertheless, the double mutant was far more sensitive to UV than either the res-1 or the recB21 strain. When compared with a wild-type strain, the rate of release of dimers from UV-irradiated DNA was very slow in the recB21 res-1, but normal in the res-1 recB+ or recB21 res+ mutants. However, the ratio of dimer-to-thymine released into the acid-soluble fraction was three times higher than the wild type in recB21 res+ and recB21 res-1 and only one-tenth as high as the wild type in res-1 rec+. Alkaline sucrose gradient centrifugation revealed occurrence of single-strand incision of UV-irradiated DNA and the restitution of nicked DNA at a similar rate in the recB21 res-1 and recB21 res+ strains. Mutants uvrC showed increased amounts of nicks in their DNA with increasing incubation time after UV irradiation, although no detectable amounts of dimers were excised from UV-irradiated DNA. From these results, it is concluded that the increased sensitivity of the res-1 strain to UV light is due to a reduced ability to excise dimers from UV-irradiated DNA and that the high rate of UV-induced breakdown of DNA is not the primary cause. A possible role of uvrC gene in the excision repair is discussed.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. BOYCE R. P., HOWARD-FLANDERS P. RELEASE OF ULTRAVIOLET LIGHT-INDUCED THYMINE DIMERS FROM DNA IN E. COLI K-12. Proc Natl Acad Sci U S A. 1964 Feb;51:293–300. doi: 10.1073/pnas.51.2.293. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Baldy M. W. Repair and recombination in phage T4. II. Genes affecting UV sensitivity. Cold Spring Harb Symp Quant Biol. 1968;33:333–338. doi: 10.1101/sqb.1968.033.01.038. [DOI] [PubMed] [Google Scholar]
  3. Barbour S. D., Clark A. J. Biochemical and genetic studies of recombination proficiency in Escherichia coli. I. Enzymatic activity associated with recB+ and recC+ genes. Proc Natl Acad Sci U S A. 1970 Apr;65(4):955–961. doi: 10.1073/pnas.65.4.955. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bazill G. W., Hall R., Gross J. D. DNA synthesis in lysates of RecB- and Rec+ E. coli cells. Nat New Biol. 1971 Oct 27;233(43):281–283. doi: 10.1038/newbio233281a0. [DOI] [PubMed] [Google Scholar]
  5. Boyle J. M., Paterson M. C., Setlow R. B. Excision-repair properties of an Escherichia coli mutant deficient in DNA polymerase. Nature. 1970 May 23;226(5247):708–710. doi: 10.1038/226708a0. [DOI] [PubMed] [Google Scholar]
  6. De Lucia P., Cairns J. Isolation of an E. coli strain with a mutation affecting DNA polymerase. Nature. 1969 Dec 20;224(5225):1164–1166. doi: 10.1038/2241164a0. [DOI] [PubMed] [Google Scholar]
  7. Emmerson P. T. Recombination deficient mutants of Escherichia coli K12 that map between thy A and argA. Genetics. 1968 Sep;60(1):19–30. doi: 10.1093/genetics/60.1.19. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Gellert M., Bullock M. L. DNA ligase mutants of Escherichia coli. Proc Natl Acad Sci U S A. 1970 Nov;67(3):1580–1587. doi: 10.1073/pnas.67.3.1580. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Goldmark P. J., Linn S. An endonuclease activity from Escherichia coli absent from certain rec- strains. Proc Natl Acad Sci U S A. 1970 Sep;67(1):434–441. doi: 10.1073/pnas.67.1.434. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Gross J. D., Grunstein J., Witkin E. M. Inviability of recA- derivatives of the DNA polymerase mutant of De Lucia and Cairns. J Mol Biol. 1971 Jun 14;58(2):631–634. doi: 10.1016/0022-2836(71)90377-9. [DOI] [PubMed] [Google Scholar]
  11. Gross J., Gross M. Genetic analysis of an E. coli strain with a mutation affecting DNA polymerase. Nature. 1969 Dec 20;224(5225):1166–1168. doi: 10.1038/2241166a0. [DOI] [PubMed] [Google Scholar]
  12. Howard-Flanders P., Boyce R. P. DNA repair and genetic recombination: studies on mutants of Escherichia coli defective in these processes. Radiat Res. 1966;(Suppl):156+–156+. [PubMed] [Google Scholar]
  13. Howard-Flanders P., Boyce R. P., Theriot L. Three loci in Escherichia coli K-12 that control the excision of pyrimidine dimers and certain other mutagen products from DNA. Genetics. 1966 Jun;53(6):1119–1136. doi: 10.1093/genetics/53.6.1119. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Kapp D. S., Smith K. C. Chemical nature of chain breaks produced in DNA by x-irradiation in vitro. Radiat Res. 1970 Apr;42(1):34–49. [PubMed] [Google Scholar]
  15. Kato T., Kondo S. Genetic and molecular characteristics of X-ray-sensitive mutants of Escherichia coli defective in repair synthesis. J Bacteriol. 1970 Nov;104(2):871–881. doi: 10.1128/jb.104.2.871-881.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Kato T., Kondo S. Two types of x-ray-sensitive mutants of Escherichia coli B: their phenotypic characters compared with UV-sensitive mutants. Mutat Res. 1967 May-Jun;4(3):253–263. doi: 10.1016/0027-5107(67)90020-6. [DOI] [PubMed] [Google Scholar]
  17. Kondo S., Ichikawa H., Iwo K., Kato T. Base-change mutagenesis and prophage induction in strains of Escherichia coli with different DNA repair capacities. Genetics. 1970 Oct;66(2):187–217. doi: 10.1093/genetics/66.2.187. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Kondo S., Kato T. Action spectra for photoreactivation of killing and mutation to prototrophy in U.V.-sensitive strains of Escherichia Coli possessing and lacking photoreactivating enzyme. Photochem Photobiol. 1966 Nov-Dec;5(11):827–837. doi: 10.1111/j.1751-1097.1966.tb05929.x. [DOI] [PubMed] [Google Scholar]
  19. Kushner S. R., Kaplan J. C., Ono H., Grossman L. Enzymatic repair of deoxyribonucleic acid. IV. Mechanism of photoproduct excision. Biochemistry. 1971 Aug 31;10(18):3325–3334. doi: 10.1021/bi00794a002. [DOI] [PubMed] [Google Scholar]
  20. Masamune Y., Fleischman R. A., Richardson C. C. Enzymatic removal and replacement of nucleotides at single strand breaks in deoxyribonucleic acid. J Biol Chem. 1971 Apr 25;246(8):2680–2691. [PubMed] [Google Scholar]
  21. McGrath R. A., Williams R. W. Reconstruction in vivo of irradiated Escherichia coli deoxyribonucleic acid; the rejoining of broken pieces. Nature. 1966 Oct 29;212(5061):534–535. doi: 10.1038/212534a0. [DOI] [PubMed] [Google Scholar]
  22. Monk M., Kinross J. Conditional lethality of recA and recB derivatives of a strain of Escherichia coli K-12 with a temperature-sensitive deoxyribonucleic acid polymerase I. J Bacteriol. 1972 Mar;109(3):971–978. doi: 10.1128/jb.109.3.971-978.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Monk M., Peacey M., Gross J. D. Repair of damage induced by ultraviolet light in DNA polymerase-defective Escherichia coli cells. J Mol Biol. 1971 Jun 14;58(2):623–630. doi: 10.1016/0022-2836(71)90376-7. [DOI] [PubMed] [Google Scholar]
  24. Ogawa H. Genetic locations of uvrD and pol genes of E. coli. Mol Gen Genet. 1970;108(4):378–381. doi: 10.1007/BF00267777. [DOI] [PubMed] [Google Scholar]
  25. Ogawa H., Shimada K., Tomizawa J. Studies on radiation-sensitive mutants of E. coli. I. Mutants defective in the repair synthesis. Mol Gen Genet. 1968 May 3;101(3):227–244. doi: 10.1007/BF00271625. [DOI] [PubMed] [Google Scholar]
  26. Oishi M. An ATP-dependent deoxyribonuclease from Escherichia coli with a possible role in genetic recombination. Proc Natl Acad Sci U S A. 1969 Dec;64(4):1292–1299. doi: 10.1073/pnas.64.4.1292. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Okazaki R., Arisawa M., Sugino A. Slow joining of newly replicated DNA chains in DNA polymerase I-deficient Escherichia coli mutants. Proc Natl Acad Sci U S A. 1971 Dec;68(12):2954–2957. doi: 10.1073/pnas.68.12.2954. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Paterson M. C., Boyle J. M., Setlow R. B. Ultraviolet- and X-ray-induced responses of a deoxyribonucleic acid polymerase-deficient mutant of Escherichia coli. J Bacteriol. 1971 Jul;107(1):61–67. doi: 10.1128/jb.107.1.61-67.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Pauling C., Hamm L. Properties of a temperature-sensitive radiation-sensitive mutant of Escherichia coli. Proc Natl Acad Sci U S A. 1968 Aug;60(4):1495–1502. doi: 10.1073/pnas.60.4.1495. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. SETLOW R. B., CARRIER W. L. THE DISAPPEARANCE OF THYMINE DIMERS FROM DNA: AN ERROR-CORRECTING MECHANISM. Proc Natl Acad Sci U S A. 1964 Feb;51:226–231. doi: 10.1073/pnas.51.2.226. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. SETLOW R. B. PHYSICAL CHANGES AND MUTAGENESIS. J Cell Physiol. 1964 Oct;64:SUPPL 1–1:68. [PubMed] [Google Scholar]
  32. Sekiguchi M., Yasuda S., Okubo S., Nakayama H., Shimada K., Takagi Y. Mechanism of repair of DNA in bacteriophage. I. Excision of pyrimidine dimers from ultraviolet-irradiated DNA by an extract of T4-infected cells. J Mol Biol. 1970 Jan 28;47(2):231–242. doi: 10.1016/0022-2836(70)90342-6. [DOI] [PubMed] [Google Scholar]
  33. Town C. D., Smith K. C., Kaplan H. S. DNA polymerase required for rapid repair of x-ray--induced DNA strand breaks in vivo. Science. 1971 May 21;172(3985):851–854. doi: 10.1126/science.172.3985.851. [DOI] [PubMed] [Google Scholar]
  34. WITKIN E. M. PHOTOREVERSAL AND "DARK REPAIR" OF MUTATIONS TO PROTOTROPHY INDUCED BY ULTRAVIOLET LIGHT IN PHOTOREACTIVABLE AND NON-PHOTOREACTIVABLE STRAINS OF ESCHERICHIA COLI. Mutat Res. 1964 May;106:22–36. doi: 10.1016/0027-5107(64)90049-1. [DOI] [PubMed] [Google Scholar]
  35. Willetts N. S., Clark A. J. Characteristics of some multiply recombination-deficient strains of Escherichia coli. J Bacteriol. 1969 Oct;100(1):231–239. doi: 10.1128/jb.100.1.231-239.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Wright M., Buttin G., Hurwitz J. The isolation and characterization from Escherichia coli of an adenosine triphosphate-dependent deoxyribonuclease directed by rec B, C genes. J Biol Chem. 1971 Nov;246(21):6543–6555. [PubMed] [Google Scholar]
  37. Yasuda S., Sekiguchi M. T4 endonuclease involved in repair of DNA. Proc Natl Acad Sci U S A. 1970 Dec;67(4):1839–1845. doi: 10.1073/pnas.67.4.1839. [DOI] [PMC free article] [PubMed] [Google Scholar]

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