Skip to main content
Nucleic Acids Research logoLink to Nucleic Acids Research
. 1985 Mar 11;13(5):1483–1492. doi: 10.1093/nar/13.5.1483

Stimulation of the UvrABC enzyme-catalyzed repair reactions by the UvrD protein (DNA helicase II).

K Kumura, M Sekiguchi, A L Steinum, E Seeberg
PMCID: PMC341090  PMID: 2987825

Abstract

An in vitro assay system was constructed using highly purified preparations of UvrA, UvrB, UvrC, UvrD proteins and DNA polymerase I, the objective being to analyse the role of UvrD protein in excision repair of UV-induced DNA damage. UvrABC enzyme-initiated repair synthesis was greatly enhanced by the addition of UvrD protein to the reaction mixture. Further analysis revealed that UvrD protein stimulated introduction of strand breaks in irradiated DNA by UvrABC enzyme but had no effect on the DNA polymerase I reaction. Thus, the site of action of UvrD protein is probably at the incision-excision step and not in later steps in excision repair.

Full text

PDF
1483

Images in this article

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. Hanawalt P. C., Cooper P. K., Ganesan A. K., Smith C. A. DNA repair in bacteria and mammalian cells. Annu Rev Biochem. 1979;48:783–836. doi: 10.1146/annurev.bi.48.070179.004031. [DOI] [PubMed] [Google Scholar]
  3. Hickson I. D., Arthur H. M., Bramhill D., Emmerson P. T. The E. coli uvrD gene product is DNA helicase II. Mol Gen Genet. 1983;190(2):265–270. doi: 10.1007/BF00330649. [DOI] [PubMed] [Google Scholar]
  4. Kuemmerle N. B., Ley R. D., Masker W. E. The effect of mutations in the uvrD cistron of Escherichia coli on repair resynthesis. Mutat Res. 1982 Jun;94(2):285–297. doi: 10.1016/0027-5107(82)90292-5. [DOI] [PubMed] [Google Scholar]
  5. Kuemmerle N. B., Masker W. E. An in vitro complementation assay for the Escherichia coli uvrD gene product. Nucleic Acids Res. 1983 Apr 11;11(7):2193–2204. doi: 10.1093/nar/11.7.2193. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Kuemmerle N. B., Masker W. E. Effect of the uvrD mutation on excision repair. J Bacteriol. 1980 May;142(2):535–546. doi: 10.1128/jb.142.2.535-546.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Kumura K., Sekiguchi M. Identification of the uvrD gene product of Escherichia coli as DNA helicase II and its induction by DNA-damaging agents. J Biol Chem. 1984 Feb 10;259(3):1560–1565. [PubMed] [Google Scholar]
  8. Maples V. F., Kushner S. R. DNA repair in Escherichia coli: identification of the uvrD gene product. Proc Natl Acad Sci U S A. 1982 Sep;79(18):5616–5620. doi: 10.1073/pnas.79.18.5616. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Maxam A. M., Gilbert W. Sequencing end-labeled DNA with base-specific chemical cleavages. Methods Enzymol. 1980;65(1):499–560. doi: 10.1016/s0076-6879(80)65059-9. [DOI] [PubMed] [Google Scholar]
  10. Oeda K., Horiuchi T., Sekiguchi M. The uvrD gene of E. coli encodes a DNA-dependent ATPase. Nature. 1982 Jul 1;298(5869):98–100. doi: 10.1038/298098a0. [DOI] [PubMed] [Google Scholar]
  11. 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]
  12. 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]
  13. 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]
  14. Sancar A., Rupp W. D. A novel repair enzyme: UVRABC excision nuclease of Escherichia coli cuts a DNA strand on both sides of the damaged region. Cell. 1983 May;33(1):249–260. doi: 10.1016/0092-8674(83)90354-9. [DOI] [PubMed] [Google Scholar]
  15. Seeberg E. Reconstitution of an Escherichia coli repair endonuclease activity from the separated uvrA+ and uvrB+/uvrC+ gene products. Proc Natl Acad Sci U S A. 1978 Jun;75(6):2569–2573. doi: 10.1073/pnas.75.6.2569. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Seeberg E., Steinum A. L. Purification and properties of the uvrA protein from Escherichia coli. Proc Natl Acad Sci U S A. 1982 Feb;79(4):988–992. doi: 10.1073/pnas.79.4.988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Shimada K., Ogawa H., Tomizawa J. Studies on radiation-sensitive mutants of E. coli. II. Breakage and repair of ultraviolet irradiated intracellular DNA of phage lambda. Mol Gen Genet. 1968 May 3;101(3):245–256. doi: 10.1007/BF00271626. [DOI] [PubMed] [Google Scholar]
  18. Sinzinis B. I., Smirnov G. B., Saenko A. A. Repair deficiency in Escherichia coli UV-sensitive mutator strain uvr502. Biochem Biophys Res Commun. 1973 Jul 2;53(1):309–316. doi: 10.1016/0006-291x(73)91435-6. [DOI] [PubMed] [Google Scholar]
  19. Taucher-Scholz G., Hoffmann-Berling H. Identification of the gene for DNA helicase II of Escherichia coli. Eur J Biochem. 1983 Dec 15;137(3):573–580. doi: 10.1111/j.1432-1033.1983.tb07864.x. [DOI] [PubMed] [Google Scholar]
  20. Van Sluis C. A., Mattern I. E., Paterson M. C. Properties of uvrE mutants of Escherichia coli K12. I. Effects of UV irradiation on DNA metabolism. Mutat Res. 1974 Dec;25(3):273–279. doi: 10.1016/0027-5107(74)90055-4. [DOI] [PubMed] [Google Scholar]
  21. Yeung A. T., Mattes W. B., Oh E. Y., Grossman L. Enzymatic properties of purified Escherichia coli uvrABC proteins. Proc Natl Acad Sci U S A. 1983 Oct;80(20):6157–6161. doi: 10.1073/pnas.80.20.6157. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Nucleic Acids Research are provided here courtesy of Oxford University Press

RESOURCES