Skip to main content
PMC home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1986 Aug;83(15):5607–5610. doi: 10.1073/pnas.83.15.5607

Suppression of human DNA alkylation-repair defects by Escherichia coli DNA-repair genes.

L Samson, B Derfler, E A Waldstein
PMCID: PMC386337  PMID: 3526337

Abstract

The ada-alkB operon protects Escherichia coli against the effects of many alkylating agents. We have subcloned it into the pSV2 mammalian expression vector to yield pSV2ada-alkB, and this plasmid has been introduced into Mer- HeLa S3 cells, which are extremely sensitive to killing and induction of sister chromatid exchange by alkylating agents. One transformant (the S3-9 cell line) has several integrated copies of pSV2ada-alkB and was found to express a very high level of the ada gene product, the 39-kDa O6-methylguanine-DNA methyltransferase. S3-9 cells were found to have become resistant to killing and induction of sister chromatid exchange by two alkylating agents, N-methyl-N'-nitro-N-nitrosoguanidine and N,N'-bis(2-chloroethyl)-N-nitro-sourea. This shows that bacterial DNA alkylation-repair genes are able to suppress the alkylation-repair defects in human Mer- cells.

Full text

PDF

Images in this article

5609Image
on p.5609
5609Image
on p.5609
5609Image
on p.5609
5609Image
on p.5609

Selected References

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

  1. Baker R. M., Van Voorhis W. C., Spencer L. A. HeLa cell variants that differ in sensitivity to monofunctional alkylating agents, with independence of cytotoxic and mutagenic responses. Proc Natl Acad Sci U S A. 1979 Oct;76(10):5249–5253. doi: 10.1073/pnas.76.10.5249. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Brennand J., Margison G. P. Expression of the E. coli O6-methylguanine-methylphosphotriester methyltransferase gene in mammalian cells. Carcinogenesis. 1986 Jan;7(1):185–188. doi: 10.1093/carcin/7.1.185. [DOI] [PubMed] [Google Scholar]
  3. Carrano A. V., Thompson L. H., Lindl P. A., Minkler J. L. Sister chromatid exchange as an indicator of mutagenesis. Nature. 1978 Feb 9;271(5645):551–553. doi: 10.1038/271551a0. [DOI] [PubMed] [Google Scholar]
  4. Day R. S., 3rd, Ziolkowski C. H., Scudiero D. A., Meyer S. A., Lubiniecki A. S., Girardi A. J., Galloway S. M., Bynum G. D. Defective repair of alkylated DNA by human tumour and SV40-transformed human cell strains. Nature. 1980 Dec 25;288(5792):724–727. doi: 10.1038/288724a0. [DOI] [PubMed] [Google Scholar]
  5. Demple B., Jacobsson A., Olsson M., Robins P., Lindahl T. Repair of alkylated DNA in Escherichia coli. Physical properties of O6-methylguanine-DNA methyltransferase. J Biol Chem. 1982 Nov 25;257(22):13776–13780. [PubMed] [Google Scholar]
  6. Ding R., Ghosh K., Eastman A., Bresnick E. DNA-mediated transfer and expression of a human DNA repair gene that demethylates O6-methylguanine. Mol Cell Biol. 1985 Nov;5(11):3293–3296. doi: 10.1128/mcb.5.11.3293. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Domoradzki J., Pegg A. E., Dolan M. E., Maher V. M., McCormick J. J. Depletion of O6-methylguanine-DNA-methyltransferase in human fibroblasts increases the mutagenic response to N-methyl-N'-nitro-N-nitrosoguanidine. Carcinogenesis. 1985 Dec;6(12):1823–1826. doi: 10.1093/carcin/6.12.1823. [DOI] [PubMed] [Google Scholar]
  8. Evensen G., Seeberg E. Adaptation to alkylation resistance involves the induction of a DNA glycosylase. Nature. 1982 Apr 22;296(5859):773–775. doi: 10.1038/296773a0. [DOI] [PubMed] [Google Scholar]
  9. Foote R. S., Pal B. C., Mitra S. Quantitation of O6-methylguanine-DNA methyltransferase in HeLa cells. Mutat Res. 1983 Mar;119(3):221–228. doi: 10.1016/0165-7992(83)90164-1. [DOI] [PubMed] [Google Scholar]
  10. Karran P., Hjelmgren T., Lindahl T. Induction of a DNA glycosylase for N-methylated purines is part of the adaptive response to alkylating agents. Nature. 1982 Apr 22;296(5859):770–773. doi: 10.1038/296770a0. [DOI] [PubMed] [Google Scholar]
  11. Karran P., Lindahl T., Griffin B. Adaptive response to alkylating agents involves alteration in situ of O6-methylguanine residues in DNA. Nature. 1979 Jul 5;280(5717):76–77. doi: 10.1038/280076a0. [DOI] [PubMed] [Google Scholar]
  12. Karran P., Lindahl T., Ofsteng I., Evensen G. B., Seeberg E. Escherichia coli mutants deficient in 3-methyladenine-DNA glycosylase. J Mol Biol. 1980 Jun 15;140(1):101–127. doi: 10.1016/0022-2836(80)90358-7. [DOI] [PubMed] [Google Scholar]
  13. Karran P. Possible depletion of a DNA repair enzyme in human lymphoma cells by subversive repair. Proc Natl Acad Sci U S A. 1985 Aug;82(16):5285–5289. doi: 10.1073/pnas.82.16.5285. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Karran P., Williams S. A. The cytotoxic and mutagenic effects of alkylating agents on human lymphoid cells are caused by different DNA lesions. Carcinogenesis. 1985 May;6(5):789–792. doi: 10.1093/carcin/6.5.789. [DOI] [PubMed] [Google Scholar]
  15. Kataoka H., Sekiguchi M. Molecular cloning and characterization of the alkB gene of Escherichia coli. Mol Gen Genet. 1985;198(2):263–269. doi: 10.1007/BF00383004. [DOI] [PubMed] [Google Scholar]
  16. Kataoka H., Yamamoto Y., Sekiguchi M. A new gene (alkB) of Escherichia coli that controls sensitivity to methyl methane sulfonate. J Bacteriol. 1983 Mar;153(3):1301–1307. doi: 10.1128/jb.153.3.1301-1307.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Lemotte P. K., Walker G. C. Induction and autoregulation of ada, a positively acting element regulating the response of Escherichia coli K-12 to methylating agents. J Bacteriol. 1985 Mar;161(3):888–895. doi: 10.1128/jb.161.3.888-895.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Loechler E. L., Green C. L., Essigmann J. M. In vivo mutagenesis by O6-methylguanine built into a unique site in a viral genome. Proc Natl Acad Sci U S A. 1984 Oct;81(20):6271–6275. doi: 10.1073/pnas.81.20.6271. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Margison G. P., Cooper D. P., Brennand J. Cloning of the E. coli O6-methylguanine and methylphosphotriester methyltransferase gene using a functional DNA repair assay. Nucleic Acids Res. 1985 Mar 25;13(6):1939–1952. doi: 10.1093/nar/13.6.1939. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. McCarthy T. V., Lindahl T. Methyl phosphotriesters in alkylated DNA are repaired by the Ada regulatory protein of E. coli. Nucleic Acids Res. 1985 Apr 25;13(8):2683–2698. doi: 10.1093/nar/13.8.2683. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Olsson M., Lindahl T. Repair of alkylated DNA in Escherichia coli. Methyl group transfer from O6-methylguanine to a protein cysteine residue. J Biol Chem. 1980 Nov 25;255(22):10569–10571. [PubMed] [Google Scholar]
  22. Perry P., Wolff S. New Giemsa method for the differential staining of sister chromatids. Nature. 1974 Sep 13;251(5471):156–158. doi: 10.1038/251156a0. [DOI] [PubMed] [Google Scholar]
  23. Robins P., Harris A. L., Goldsmith I., Lindahl T. Cross-linking of DNA induced by chloroethylnitrosourea is presented by O6-methylguanine-DNA methyltransferase. Nucleic Acids Res. 1983 Nov 25;11(22):7743–7758. doi: 10.1093/nar/11.22.7743. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Schendel P. F., Robins P. E. Repair of O6-methylguanine in adapted Escherichia coli. Proc Natl Acad Sci U S A. 1978 Dec;75(12):6017–6020. doi: 10.1073/pnas.75.12.6017. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Scudiero D. A., Meyer S. A., Clatterbuck B. E., Mattern M. R., Ziolkowski C. H., Day R. S., 3rd Sensitivity of human cell strains having different abilities to repair O6-methylguanine in DNA to inactivation by alkylating agents including chloroethylnitrosoureas. Cancer Res. 1984 Jun;44(6):2467–2474. [PubMed] [Google Scholar]
  26. Sklar R., Strauss B. Removal of O6-methylguanine from DNA of normal and xeroderma pigmentosum-derived lymphoblastoid lines. Nature. 1981 Jan 29;289(5796):417–420. doi: 10.1038/289417a0. [DOI] [PubMed] [Google Scholar]
  27. Southern P. J., Berg P. Transformation of mammalian cells to antibiotic resistance with a bacterial gene under control of the SV40 early region promoter. J Mol Appl Genet. 1982;1(4):327–341. [PubMed] [Google Scholar]
  28. Teo I., Sedgwick B., Demple B., Li B., Lindahl T. Induction of resistance to alkylating agents in E. coli: the ada+ gene product serves both as a regulatory protein and as an enzyme for repair of mutagenic damage. EMBO J. 1984 Sep;3(9):2151–2157. doi: 10.1002/j.1460-2075.1984.tb02105.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Valerie K., de Riel J. K., Henderson E. E. Genetic complementation of UV-induced DNA repair in Chinese hamster ovary cells by the denV gene of phage T4. Proc Natl Acad Sci U S A. 1985 Nov;82(22):7656–7660. doi: 10.1073/pnas.82.22.7656. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES