Abstract
The E.coli ada gene protein coding region has been ligated into an extrachromosomally replicating yeast expression vector downstream of the yeast alcohol dehydrogenase gene promoter region to produce pADH06C. The yeast strains SX46A, 7799-4B and VV-6 are deficient in endogenous O6-alkylguanine-DNA-alkyltransferase and transformation of these strains with this shuttle vector resulted in the expression of 1730, 1260 and 374 fmoles ada-encoded ATase/mg protein in stationary phase yeast: transformation with the parent vector had no effect on endogenous ATase activity which remained less than 2 fm/mg. In comparison with parent vector transformed yeast, all of the pADH06C-transformed strains showed an increase in the resistance to the toxic effects of N-methyl-N'-nitro-N-nitrosoguanidine (MNNG). In addition, 7799-4B and VV-6 were more resistant to the mutagenic effects of this agent. These results indicate that the toxic and mutagenic effects of MNNG in yeast are mediated, at least in part, by DNA lesions than can be repaired by the E.coli ada gene product.
Full text
PDF




Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Ammerer G. Expression of genes in yeast using the ADCI promoter. Methods Enzymol. 1983;101:192–201. doi: 10.1016/0076-6879(83)01014-9. [DOI] [PubMed] [Google Scholar]
- Brennand J., Margison G. P. Expression in mammalian cells of a truncated Escherichia coli gene coding for O6-alkylguanine alkyltransferase reduces the toxic effects of alkylating agents. Carcinogenesis. 1986 Dec;7(12):2081–2084. doi: 10.1093/carcin/7.12.2081. [DOI] [PubMed] [Google Scholar]
- 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]
- Brennand J., Margison G. P. Reduction of the toxicity and mutagenicity of alkylating agents in mammalian cells harboring the Escherichia coli alkyltransferase gene. Proc Natl Acad Sci U S A. 1986 Sep;83(17):6292–6296. doi: 10.1073/pnas.83.17.6292. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Chenevert J. M., Naumovski L., Schultz R. A., Friedberg E. C. Partial complementation of the UV sensitivity of E. coli and yeast excision repair mutants by the cloned denV gene of bacteriophage T4. Mol Gen Genet. 1986 Apr;203(1):163–171. doi: 10.1007/BF00330398. [DOI] [PubMed] [Google Scholar]
- Fox M., Brennand J., Margison G. P. Protection of Chinese hamster cells against the cytotoxic and mutagenic effects of alkylating agents by transfection of the Escherichia coli alkyltransferase gene and a truncated derivative. Mutagenesis. 1987 Nov;2(6):491–496. doi: 10.1093/mutage/2.6.491. [DOI] [PubMed] [Google Scholar]
- Hadden C. T., Foote R. S., Mitra S. Adaptive response of Bacillus subtilis to N-methyl-N'-nitro-N-nitrosoguanidine. J Bacteriol. 1983 Feb;153(2):756–762. doi: 10.1128/jb.153.2.756-762.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ito H., Fukuda Y., Murata K., Kimura A. Transformation of intact yeast cells treated with alkali cations. J Bacteriol. 1983 Jan;153(1):163–168. doi: 10.1128/jb.153.1.163-168.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Langeveld S. A., Yasui A., Eker A. P. Expression of an Escherichia coli phr gene in the yeast Saccharomyces cerevisiae. Mol Gen Genet. 1985;199(3):396–400. doi: 10.1007/BF00330748. [DOI] [PubMed] [Google Scholar]
- Maga J. A., McEntee K. Response of S. cerevisiae to N-methyl-N'-nitro-N-nitrosoguanidine: mutagenesis, survival and DDR gene expression. Mol Gen Genet. 1985;200(2):313–321. doi: 10.1007/BF00425442. [DOI] [PubMed] [Google Scholar]
- Margison G. P., Brennand J., Ockey C. H., O'Connor P. J. Exploring molecular mechanisms in chemically induced cancer: complementation of mammalian DNA repair defects by a prokaryotic gene. Bioessays. 1987 Apr;6(4):151–156. doi: 10.1002/bies.950060402. [DOI] [PubMed] [Google Scholar]
- McCarthy T. V., Karran P., Lindahl T. Inducible repair of O-alkylated DNA pyrimidines in Escherichia coli. EMBO J. 1984 Mar;3(3):545–550. doi: 10.1002/j.1460-2075.1984.tb01844.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Morten J. E., Margison G. P. Increased O6-alkylguanine alkyltransferase activity in Chinese hamster V79 cells following selection with chloroethylating agents. Carcinogenesis. 1988 Jan;9(1):45–49. doi: 10.1093/carcin/9.1.45. [DOI] [PubMed] [Google Scholar]
- Polakowska R., Perozzi G., Prakash L. Alkylation mutagenesis in Saccharomyces cerevisiae: lack of evidence for an adaptive response. Curr Genet. 1986;10(9):647–655. doi: 10.1007/BF00410912. [DOI] [PubMed] [Google Scholar]
- Saffhill R., Margison G. P., O'Connor P. J. Mechanisms of carcinogenesis induced by alkylating agents. Biochim Biophys Acta. 1985 Dec 17;823(2):111–145. doi: 10.1016/0304-419x(85)90009-5. [DOI] [PubMed] [Google Scholar]
- Sedgwick B. Molecular signal for induction of the adaptive response to alkylation damage in Escherichia coli. J Cell Sci Suppl. 1987;6:215–223. doi: 10.1242/jcs.1984.supplement_6.15. [DOI] [PubMed] [Google Scholar]
- Singer B., Kuśmierek J. T. Chemical mutagenesis. Annu Rev Biochem. 1982;51:655–693. doi: 10.1146/annurev.bi.51.070182.003255. [DOI] [PubMed] [Google Scholar]
- White G. R., Ockey C. H., Brennand J., Margison G. P. Chinese hamster cells harbouring the Escherichia coli O6-alkylguanine alkyltransferase gene are less susceptible to sister chromatid exchange induction and chromosome damage by methylating agents. Carcinogenesis. 1986 Dec;7(12):2077–2080. doi: 10.1093/carcin/7.12.2077. [DOI] [PubMed] [Google Scholar]