Skip to main content
NCBI home page
Nucleic Acids Research logoLink to Nucleic Acids Research
. 1990 Jan 11;18(1):17–21. doi: 10.1093/nar/18.1.17

Active site amino acid sequence of the bovine O6-methylguanine-DNA methyltransferase.

B Rydberg 1, J Hall 1, P Karran 1
PMCID: PMC330198  PMID: 2308822

Abstract

An O6-methylguanine-DNA methyltransferase has been partially purified from calf thymus by conventional biochemical techniques. The enzyme was specifically radioactively labelled at the cysteine residue of the active site and further purified by attachment to a solid support. Following digestion with trypsin, a radioactive peptide containing the active site region of the protein was purified by size fractionation, ion exchange chromatography and reverse phase HPLC. The technique yielded an essentially homogeneous oligopeptide which was subjected to amino acid sequencing. The sequence adjacent to the acceptor cysteine residue of the bovine protein exhibits striking homology to the C-terminal methyl acceptor site of the E. coli Ada protein and the proposed acceptor sites of the E. coli Ogt and the B. subtilis Dat1 proteins.

Full text

PDF
17

Selected References

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

  1. Abbott P. J., Saffhill R. DNA synthesis with methylated poly(dC-dG) templates. Evidence for a competitive nature to miscoding by O(6)-methylguanine. Biochim Biophys Acta. 1979 Mar 28;562(1):51–61. doi: 10.1016/0005-2787(79)90125-4. [DOI] [PubMed] [Google Scholar]
  2. Ather A., Ahmed Z., Riazuddin S. Adaptive response of Micrococcus luteus to alkylating chemicals. Nucleic Acids Res. 1984 Feb 24;12(4):2111–2126. doi: 10.1093/nar/12.4.2111. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. 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]
  4. 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]
  5. Demple B., Sedgwick B., Robins P., Totty N., Waterfield M. D., Lindahl T. Active site and complete sequence of the suicidal methyltransferase that counters alkylation mutagenesis. Proc Natl Acad Sci U S A. 1985 May;82(9):2688–2692. doi: 10.1073/pnas.82.9.2688. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Hall J., Kataoka H., Stephenson C., Karran P. The contribution of O6-methylguanine and methylphosphotriesters to the cytotoxicity of alkylating agents in mammalian cells. Carcinogenesis. 1988 Sep;9(9):1587–1593. doi: 10.1093/carcin/9.9.1587. [DOI] [PubMed] [Google Scholar]
  7. Harris A. L., Karran P., Lindahl T. O6-Methylguanine-DNA methyltransferase of human lymphoid cells: structural and kinetic properties and absence in repair-deficient cells. Cancer Res. 1983 Jul;43(7):3247–3252. [PubMed] [Google Scholar]
  8. Hill-Perkins M., Jones M. D., Karran P. Site-specific mutagenesis in vivo by single methylated or deaminated purine bases. Mutat Res. 1986 Sep;162(2):153–163. doi: 10.1016/0027-5107(86)90081-3. [DOI] [PubMed] [Google Scholar]
  9. Hora J. F., Eastman A., Bresnick E. O6-methylguanine methyltransferase in rat liver. Biochemistry. 1983 Aug 2;22(16):3759–3763. doi: 10.1021/bi00285a007. [DOI] [PubMed] [Google Scholar]
  10. Ishizaki K., Tsujimura T., Fujio C., Zhang Y. P., Yawata H., Nakabeppu Y., Sekiguchi M., Ikenaga M. Expression of the truncated E. coli O6-methylguanine methyltransferase gene in repair-deficient human cells and restoration of cellular resistance to alkylating agents. Mutat Res. 1987 Sep;184(2):121–128. doi: 10.1016/0167-8817(87)90068-x. [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. Lindahl T., Sedgwick B., Sekiguchi M., Nakabeppu Y. Regulation and expression of the adaptive response to alkylating agents. Annu Rev Biochem. 1988;57:133–157. doi: 10.1146/annurev.bi.57.070188.001025. [DOI] [PubMed] [Google Scholar]
  13. 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]
  14. Morohoshi F., Hayashi K., Munakata N. Bacillus subtilis gene coding for constitutive O6-methylguanine-DNA alkyltransferase. Nucleic Acids Res. 1989 Aug 25;17(16):6531–6543. doi: 10.1093/nar/17.16.6531. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Morohoshi F., Munakata N. Multiple species of Bacillus subtilis DNA alkyltransferase involved in the adaptive response to simple alkylating agents. J Bacteriol. 1987 Feb;169(2):587–592. doi: 10.1128/jb.169.2.587-592.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Pegg A. E., Wiest L., Foote R. S., Mitra S., Perry W. Purification and properties of O6-methylguanine-DNA transmethylase from rat liver. J Biol Chem. 1983 Feb 25;258(4):2327–2333. [PubMed] [Google Scholar]
  17. Potter P. M., Wilkinson M. C., Fitton J., Carr F. J., Brennand J., Cooper D. P., Margison G. P. Characterisation and nucleotide sequence of ogt, the O6-alkylguanine-DNA-alkyltransferase gene of E. coli. Nucleic Acids Res. 1987 Nov 25;15(22):9177–9193. doi: 10.1093/nar/15.22.9177. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Pósfai J., Bhagwat A. S., Pósfai G., Roberts R. J. Predictive motifs derived from cytosine methyltransferases. Nucleic Acids Res. 1989 Apr 11;17(7):2421–2435. doi: 10.1093/nar/17.7.2421. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Sedgwick B., Robins P., Totty N., Lindahl T. Functional domains and methyl acceptor sites of the Escherichia coli ada protein. J Biol Chem. 1988 Mar 25;263(9):4430–4433. [PubMed] [Google Scholar]
  20. 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]

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

RESOURCES