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. 1977 Jul;4(7):2445–2454. doi: 10.1093/nar/4.7.2445

Characterization of human enzymes specific for damaged DNA: resolution of endonuclease for irradiated DNA from an apparent N-glycosidase active on alkylated DNA.

T P Brent
PMCID: PMC342577  PMID: 198741

Abstract

An endonuclease partially purified from human lymphoblasts, and active against ultraviolet-irradiated DNA, was found to act additionally on DNA damaged by either x-radiation or methylmethanesulfonate. To determine if these activities were truly endonucleolytic, the reaction products were analyzed under conditions that prevented conversion of apurinic or apyrimidinic sites to single-strand breaks. With either ultraviolet- or x-irradiated DNA, strand breakage remained maximal, hence confirming the endonucleolytic character of the enzyme. By contrast, with DNA alkylated with methylmethanesulfonate, strand breakage was sharply reduced. Additional experiments indicated that the activity for alkylated DNA induces strand breaks only in concert with a purified endonuclease specific for apurinic sites, suggesting that it is an N-glycosidase that depurinates alkylated bases. This enzyme was separated from the endonuclease specific for irradiated DNA, by chromatography on DNA-agarose.

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Selected References

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

  1. Bacchetti S., Benne R. Purification and characterization of an endonuclease from calf thymus acting on irradiated DNA. Biochim Biophys Acta. 1975 May 16;390(3):285–297. doi: 10.1016/0005-2787(75)90349-4. [DOI] [PubMed] [Google Scholar]
  2. Braun A. G., Radman M., Grossman L. Enzymatic repair of DNA: SITES OF HYDROLYSIS BY THE Escherichia coli endonuclease specific for pyrimidine dimers (correndonuclease II). Biochemistry. 1976 Sep 7;15(18):4116–4120. doi: 10.1021/bi00663a031. [DOI] [PubMed] [Google Scholar]
  3. Brent T. P. A human endonuclease activity for gamma-irradiated DNA. Biophys J. 1973 Apr;13(4):399–401. doi: 10.1016/S0006-3495(73)85993-4. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Brent T. P. Partial purification of endonuclease activity from human lymphoblasts. Separation of activities for depurinated DNA and DNA irradiated with ultraviolet light. Biochim Biophys Acta. 1975 Oct 1;407(2):191–199. doi: 10.1016/0005-2787(75)90284-1. [DOI] [PubMed] [Google Scholar]
  5. Brent T. P. Purification and characterization of human endonucleases specific for damaged DNA. Analysis of lesions induced by ultraviolet or x-radiation. Biochim Biophys Acta. 1976 Nov 12;454(1):172–183. doi: 10.1016/0005-2787(76)90363-4. [DOI] [PubMed] [Google Scholar]
  6. Brent T. P. Repair enzyme suggested by mammalian endonuclease activity specific for ultraviolet-irradiated DNA. Nat New Biol. 1972 Oct 11;239(93):172–173. doi: 10.1038/newbio239172a0. [DOI] [PubMed] [Google Scholar]
  7. Carrier W. L., Setlow R. B. Endonuclease from Micrococcus luteus which has activity toward ultraviolet-irradiated deoxyribonucleic acid: purification and properties. J Bacteriol. 1970 Apr;102(1):178–186. doi: 10.1128/jb.102.1.178-186.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Duker N. J., Teebor G. W. Detection of different types of damage in alkylated DNA by means of human corrective endonuclease (correndonuclease). Proc Natl Acad Sci U S A. 1976 Aug;73(8):2629–2633. doi: 10.1073/pnas.73.8.2629. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Duker N. J., Teebor G. W. Different ultraviolet DNA endonuclease activity in human cells. Nature. 1975 May 1;255(5503):82–84. doi: 10.1038/255082a0. [DOI] [PubMed] [Google Scholar]
  10. Espejo R. T., Canelo E. S. Properties of bacteriophage PM2: a lipid-containing bacterial virus. Virology. 1968 Apr;34(4):738–747. doi: 10.1016/0042-6822(68)90094-9. [DOI] [PubMed] [Google Scholar]
  11. Espejo R. T., Canelo E. S., Sinsheimer R. L. DNA of bacteriophage PM2: a closed circular double-stranded molecule. Proc Natl Acad Sci U S A. 1969 Aug;63(4):1164–1168. doi: 10.1073/pnas.63.4.1164. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Friedberg E. C., Ganesan A. K., Minton K. N-Glycosidase activity in extracts of Bacillus subtilis and its inhibition after infection with bacteriophage PBS2. J Virol. 1975 Aug;16(2):315–321. doi: 10.1128/jvi.16.2.315-321.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Friedberg E. C., King J. J. Dark repair of ultraviolet-irradiated deoxyribonucleic acid by bacteriophage T4: purification and characterization of a dimer-specific phage-induced endonuclease. J Bacteriol. 1971 May;106(2):500–507. doi: 10.1128/jb.106.2.500-507.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Kaplan J. C., Kushner S. R., Grossman L. Enzymatic repair of DNA. 3. Properties of the UV-endonuclease and UV-exonuclease. Biochemistry. 1971 Aug 31;10(18):3315–3324. doi: 10.1021/bi00794a001. [DOI] [PubMed] [Google Scholar]
  15. 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]
  16. Kirtikar D. M., Goldthwait D. A. The enzymatic release of O6-methylguanine and 3-methyladenine from DNA reacted with the carcinogen N-methyl-N-nitrosourea. Proc Natl Acad Sci U S A. 1974 May;71(5):2022–2026. doi: 10.1073/pnas.71.5.2022. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Lindahl T. New class of enzymes acting on damaged DNA. Nature. 1976 Jan 1;259(5538):64–66. doi: 10.1038/259064a0. [DOI] [PubMed] [Google Scholar]
  18. Ljungquist S., Lindahl T. A mammalian endonuclease specific for apurinic sites in double-stranded deoxyribonucleic acid. I. Purification and general properties. J Biol Chem. 1974 Mar 10;249(5):1530–1535. [PubMed] [Google Scholar]
  19. Paquette Y., Crine P., Verly W. G. Properties of the endonuclease for depurinated DNA from Escherichia coli. Can J Biochem. 1972 Nov;50(11):1199–1209. doi: 10.1139/o72-163. [DOI] [PubMed] [Google Scholar]
  20. Paterson M. C., Setlow R. B. Endonucleolytic activity from Micrococcus luteus that acts on -ray-induced damage in plasmid DNA of Escherichia coli minicells. Proc Natl Acad Sci U S A. 1972 Oct;69(10):2927–2931. doi: 10.1073/pnas.69.10.2927. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Paterson M. C., Smith B. P., Lohman P. H., Anderson A. K., Fishman L. Defective excision repair of gamma-ray-damaged DNA in human (ataxia telangiectasia) fibroblasts. Nature. 1976 Apr 1;260(5550):444–447. doi: 10.1038/260444a0. [DOI] [PubMed] [Google Scholar]
  22. Sekiguchi M., Hayakawa H., Makino F., Tanaka K., Okada Y. A human enzyme that liberates uracil from DNA. Biochem Biophys Res Commun. 1976 Nov 22;73(2):293–299. doi: 10.1016/0006-291x(76)90706-3. [DOI] [PubMed] [Google Scholar]
  23. Walker I. G., Ewart D. F. The nature of single-strand breaks in DNA following treatment of L-cells with methylating agents. Mutat Res. 1973 Sep;19(3):331–341. doi: 10.1016/0027-5107(73)90234-0. [DOI] [PubMed] [Google Scholar]
  24. 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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