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Hoffmann, R. A wiki for the life sciences where authorship matters. Nature Genetics (2008)

Molecular analysis of O6-substituted guanine-induced mutagenesis of ras oncogenes.

We have designed an Ha-ras/ thymidine kinase ( TK) cassette that permits the incorporation of chemically synthesized adducts within specific domains of the rat Ha-ras protooncogene. This cassette has been used to evaluate the mutagenicity of O6-substituted guanine residues, including O6-methylguanine and O6-benzylguanine, incorporated within the 12th codon of this locus. Mutations were monitored by the ability of these modified Ha-ras DNAs to transform Rat4 TK-cells. Our results indicate that both types of O6-substituted guanines are substantially mutagenic, although the methyl substituent induced a 2-fold higher percentage of transformed Rat4 TK+ colonies than its bulkier benzyl analogue. Interestingly, the mutagenicity of both O6-substituted guanines was found to be independent of their relative position within codon 12, therefore suggesting that the specific activation of Ha-ras oncogenes by GGA----GAA mutations in tumors induced by methylating carcinogens might be due to differences in the accessibility of these guanine residues to the carcinogen rather than to a differential rate of repair. Molecular analysis of the mutations induced by these O6-substituted guanines indicated that O6-methylguanine exclusively induced G----A transitions. In contrast, O6-benzylguanine produced G----C and G----T transversions in addition to G----A transitions. These results suggest that O6-methylguanine and its bulkier analogue O6-benzylguanine may induce mutagenesis by different mechanisms.[1]


  1. Molecular analysis of O6-substituted guanine-induced mutagenesis of ras oncogenes. Mitra, G., Pauly, G.T., Kumar, R., Pei, G.K., Hughes, S.H., Moschel, R.C., Barbacid, M. Proc. Natl. Acad. Sci. U.S.A. (1989) [Pubmed]
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