Skip to main content
PMC home page
Molecular and Cellular Biology logoLink to Molecular and Cellular Biology
. 1996 Jul;16(7):3714–3719. doi: 10.1128/mcb.16.7.3714

Site-specific excision repair of 1-nitrosopyrene-induced DNA adducts at the nucleotide level in the HPRT gene of human fibroblasts: effect of adduct conformation on the pattern of site-specific repair.

D Wei 1, V M Maher 1, J J McCormick 1
PMCID: PMC231367  PMID: 8668188

Abstract

Studies showing that different types of DNA adducts are repaired in human cells at different rates suggest that DNA adduct conformation is the major determinant of the rate of nucleotide excision repair. However, recent studies of repair of cyclobutane pyrimidine dimers or benzo[a]pyrene diol epoxide (BPDE)-induced adducts at the nucleotide level in DNA of normal human fibroblasts indicate that the rate of repair of the same adduct at different nucleotide positions can vary up to 10-fold, suggesting an important role for local DNA conformation. To see if site-specific DNA repair is a common phenomenon for bulky DNA adducts, we determined the rate of repair of 1-nitrosopyrene (1-NOP)-induced adducts in exon 3 of the hypoxanthine phosphoribosyltransferase gene at the nucleotide level using ligation-mediated PCR. To distinguish between the contributions of adduct conformation and local DNA conformation to the rate of repair, we compared the results obtained with 1-NOP with those we obtained previously using BPDE. The principal DNA adduct formed by either agent involves guanine. We found that rates of repair of 1-NOP-induced adducts also varied significantly at the nucleotide level, but the pattern of site-specific repair differed from that of BPDE-induced adducts at the same guanine positions in the same region of DNA. The average rate of excision repair of 1-NOP adducts in exon 3 was two to three times faster than that of BPDE adducts, but at particular nucleotides the rate was slower or faster than that of BPDE adducts or, in some cases, equal to that of BPDE adducts. These results indicate that the contribution of the local DNA conformation to the rate of repair at a particular nucleotide position depends upon the specific DNA adduct involved. However, the data also indicate that the conformation of the DNA adduct is not the only factor contributing to the rate of repair at different nucleotide positions. Instead, the rate of repair at a particular nucleotide position depends on the interaction between the specific adduct conformation and the local DNA conformation at that nucleotide.

Full Text

The Full Text of this article is available as a PDF (360.2 KB).

Selected References

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

  1. Beland F. A., Ribovich M., Howard P. C., Heflich R. H., Kurian P., Milo G. E. Cytotoxicity, cellular transformation and DNA adducts in normal human diploid fibroblasts exposed to 1-nitrosopyrene, a reduced derivative of the environmental contaminant, 1-nitropyrene. Carcinogenesis. 1986 Aug;7(8):1279–1283. doi: 10.1093/carcin/7.8.1279. [DOI] [PubMed] [Google Scholar]
  2. Bohr V. A., Hanawalt P. C. DNA repair in genes. Pharmacol Ther. 1988;38(3):305–319. doi: 10.1016/0163-7258(88)90008-3. [DOI] [PubMed] [Google Scholar]
  3. Bohr V. A., Smith C. A., Okumoto D. S., Hanawalt P. C. DNA repair in an active gene: removal of pyrimidine dimers from the DHFR gene of CHO cells is much more efficient than in the genome overall. Cell. 1985 Feb;40(2):359–369. doi: 10.1016/0092-8674(85)90150-3. [DOI] [PubMed] [Google Scholar]
  4. Chen R. H., Maher V. M., Brouwer J., van de Putte P., McCormick J. J. Preferential repair and strand-specific repair of benzo[a]pyrene diol epoxide adducts in the HPRT gene of diploid human fibroblasts. Proc Natl Acad Sci U S A. 1992 Jun 15;89(12):5413–5417. doi: 10.1073/pnas.89.12.5413. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Cosman M., de los Santos C., Fiala R., Hingerty B. E., Singh S. B., Ibanez V., Margulis L. A., Live D., Geacintov N. E., Broyde S. Solution conformation of the major adduct between the carcinogen (+)-anti-benzo[a]pyrene diol epoxide and DNA. Proc Natl Acad Sci U S A. 1992 Mar 1;89(5):1914–1918. doi: 10.1073/pnas.89.5.1914. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Gao S., Drouin R., Holmquist G. P. DNA repair rates mapped along the human PGK1 gene at nucleotide resolution. Science. 1994 Mar 11;263(5152):1438–1440. doi: 10.1126/science.8128226. [DOI] [PubMed] [Google Scholar]
  7. Heflich R. H., Hazard R. M., Lommel L., Scribner J. D., Maher V. M., McCormick J. J. A comparison of the DNA binding, cytotoxicity and repair synthesis induced in human fibroblasts by reactive derivatives of aromatic amide carcinogens. Chem Biol Interact. 1980 Jan;29(1):43–56. doi: 10.1016/0009-2797(80)90085-x. [DOI] [PubMed] [Google Scholar]
  8. Huang J. C., Hsu D. S., Kazantsev A., Sancar A. Substrate spectrum of human excinuclease: repair of abasic sites, methylated bases, mismatches, and bulky adducts. Proc Natl Acad Sci U S A. 1994 Dec 6;91(25):12213–12217. doi: 10.1073/pnas.91.25.12213. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Lin J. J., Sancar A. (A)BC excinuclease: the Escherichia coli nucleotide excision repair enzyme. Mol Microbiol. 1992 Aug;6(16):2219–2224. doi: 10.1111/j.1365-2958.1992.tb01398.x. [DOI] [PubMed] [Google Scholar]
  10. 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]
  11. McGregor W. G., Mah M. C., Chen R. W., Maher V. M., McCormick J. J. Lack of correlation between degree of interference with transcription and rate of strand specific repair in the HPRT gene of diploid human fibroblasts. J Biol Chem. 1995 Nov 10;270(45):27222–27227. doi: 10.1074/jbc.270.45.27222. [DOI] [PubMed] [Google Scholar]
  12. Mellon I., Hanawalt P. C. Induction of the Escherichia coli lactose operon selectively increases repair of its transcribed DNA strand. Nature. 1989 Nov 2;342(6245):95–98. doi: 10.1038/342095a0. [DOI] [PubMed] [Google Scholar]
  13. Mellon I., Spivak G., Hanawalt P. C. Selective removal of transcription-blocking DNA damage from the transcribed strand of the mammalian DHFR gene. Cell. 1987 Oct 23;51(2):241–249. doi: 10.1016/0092-8674(87)90151-6. [DOI] [PubMed] [Google Scholar]
  14. Nolan S. J., Vyas R. R., Hingerty B. E., Ellis S., Broyde S., Shapiro R., Basu A. K. Solution properties and computational analysis of an oligodeoxynucleotide containing N-(deoxyguanosin-8-yl)-1-aminopyrene. Carcinogenesis. 1996 Jan;17(1):133–144. doi: 10.1093/carcin/17.1.133. [DOI] [PubMed] [Google Scholar]
  15. Patton J. D., Maher V. M., McCormick J. J. Cytotoxic and mutagenic effects of 1-nitropyrene and 1-nitrosopyrene in diploid human fibroblasts. Carcinogenesis. 1986 Jan;7(1):89–93. doi: 10.1093/carcin/7.1.89. [DOI] [PubMed] [Google Scholar]
  16. Sancar A. Mechanisms of DNA excision repair. Science. 1994 Dec 23;266(5193):1954–1956. doi: 10.1126/science.7801120. [DOI] [PubMed] [Google Scholar]
  17. Sancar A., Tang M. S. Nucleotide excision repair. Photochem Photobiol. 1993 May;57(5):905–921. doi: 10.1111/j.1751-1097.1993.tb09233.x. [DOI] [PubMed] [Google Scholar]
  18. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Tornaletti S., Pfeifer G. P. Slow repair of pyrimidine dimers at p53 mutation hotspots in skin cancer. Science. 1994 Mar 11;263(5152):1436–1438. doi: 10.1126/science.8128225. [DOI] [PubMed] [Google Scholar]
  20. Tung B. S., McGregor W. G., Wang Y. C., Maher V. M., McCormick J. J. Comparison of the rate of excision of major UV photoproducts in the strands of the human HPRT gene of normal and xeroderma pigmentosum variant cells. Mutat Res. 1996 Jan 2;362(1):65–74. doi: 10.1016/0921-8777(95)00034-8. [DOI] [PubMed] [Google Scholar]
  21. Van Houten B., Masker W. E., Carrier W. L., Regan J. D. Quantitation of carcinogen-induced DNA damage and repair in human cells with the UVR ABC excision nuclease from Escherichia coli. Carcinogenesis. 1986 Jan;7(1):83–87. doi: 10.1093/carcin/7.1.83. [DOI] [PubMed] [Google Scholar]
  22. Wang Y., Parks W. C., Wigle J. C., Maher V. M., McCormick J. J. Fibroblasts from patients with inherited predisposition to retinoblastoma exhibit normal sensitivity to the mutagenic effects of ionizing radiation. Mutat Res. 1986 Oct;175(2):107–114. doi: 10.1016/0165-7992(86)90133-8. [DOI] [PubMed] [Google Scholar]
  23. Watanabe M., Maher V. M., McCormick J. J. Excision repair of UV- or benzo[a]pyrene diol epoxide-induced lesions in xeroderma pigmentosum variant cells is 'error free'. Mutat Res. 1985 Nov;146(3):285–294. doi: 10.1016/0167-8817(85)90070-7. [DOI] [PubMed] [Google Scholar]
  24. Wei D., Maher V. M., McCormick J. J. Site-specific rates of excision repair of benzo[a]pyrene diol epoxide adducts in the hypoxanthine phosphoribosyltransferase gene of human fibroblasts: correlation with mutation spectra. Proc Natl Acad Sci U S A. 1995 Mar 14;92(6):2204–2208. doi: 10.1073/pnas.92.6.2204. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Weinstein I. B., Jeffrey A. M., Jennette K. W., Blobstein S. H., Harvey R. G., Harris C., Autrup H., Kasai H., Nakanishi K. Benzo(a)pyrene diol epoxides as intermediates in nucleic acid binding in vitro and in vivo. Science. 1976 Aug 13;193(4253):592–595. doi: 10.1126/science.959820. [DOI] [PubMed] [Google Scholar]
  26. Yang J. L., Maher V. M., McCormick J. J. Kinds and spectrum of mutations induced by 1-nitrosopyrene adducts during plasmid replication in human cells. Mol Cell Biol. 1988 Aug;8(8):3364–3372. doi: 10.1128/mcb.8.8.3364. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Yang L. L., Maher V. M., McCormick J. J. Error-free excision of the cytotoxic,mutagenic N2-deoxyguanosine DNA adduct formed in human fibroblasts by (+/-)-7 beta, 8 alpha-dihydroxy-9 alpha, 10 alpha-epoxy-7,8,9,10-tetrahydrobenzo[a]pyrene. Proc Natl Acad Sci U S A. 1980 Oct;77(10):5933–5937. doi: 10.1073/pnas.77.10.5933. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Yang L. L., Maher V. M., McCormick J. J. Relationship between excision repair and the cytotoxic and mutagenic effect of the 'anti' 7,8-diol-9,10-epoxide of benzo[a]pyrene in human cells. Mutat Res. 1982 Jun;94(2):435–447. doi: 10.1016/0027-5107(82)90306-2. [DOI] [PubMed] [Google Scholar]

Articles from Molecular and Cellular Biology are provided here courtesy of Taylor & Francis

RESOURCES