Summary
The radiation-sensitive rad mutants of the yeast Saccharomyces cerevisiae exhibit a complex pattern of sensitivity to simple monofunctional alkylating agents. The RAD1, RAD2, RAD4 and RAD14 genes of the RAD3 epistasis group are implicated in the repair of ethylations to DNA. The RAD3, RAD10 and RAD16 genes of this group are not involved. The RAD4 and RAD14 genes have a particular role in repair following exposure to those ethylating agents that preferentially alkylate oxygen, but not to those that preferentially ethylate nitrogen. The RAD1 and RAD2 genes are involved in the repair of damage induced by all the ethylating agents used except EMS. The mutants in this group that are sensitive to ENU were not sensitive to MNU, suggesting that nucleotide excision operates on ethylations but not on methylations.
In the RAD6 group, the RAD6 and RAD18 genes are involved in DNA repair after exposure to all the alkylating agents tested, whereas RAD8 appears to have a role in the repair of O-alkylations but not N-alkylations. RAD9 operates in the repair of methylations and ethylations, but does not influence events after exposure to EMS. In the RAD52 group, the mutants tested were sensitive to ENU and DES. Thus some members of all three epistasis groups are involved in the repair of alkylations to DNA.
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Abbreviations
- DES:
-
diethylsulphate
- EMS:
-
ethylmethanesulphonate
- ENNG:
-
N-ethyl-N′-nitro-N-nitrosoguanidine
- ENU:
-
N-ethylnitrosourea
- MNU:
-
N-methylnitrosourea
- DMSO:
-
dimethylsulphoxide
- MMS:
-
methylmethanesulphonate
References
Ahmed Z, Laval J (1984) Enzymatic repair of O-alkylated thymidine residues in DNA: involvement of an O4-methylthymine DNA methyltransferase and an O2-methylthymine DNA glycosylase. Biochem Biophys Res Commun 120:1–8
Akhmedor AT, Kaboev OK, Bekker ML (1982) Purification and properties of two endonucleases specific for apurinic/apyrimidinic sites in DNA from Saccharomyces cerevisiae. Biochim Biophys Acta 696:163–168
Armel PR, Wallace SS (1978) Apurinic endonucleases from Saccharomyces cerevisiae. Nucleic Acids Res 5:3347–3356
Averbeck D, Moustacchi E (1975) 8-methoxypsoralen plus 365nm light effects and repair in yeast. Biochim Biophys Acta 395:393–404
Brendel M, Haynes RH (1973) Interactions among genes controlling sensitivity to radiation and alkylation in yeast. Mol Gen Genet 125:197–216
Brennand J, Margison GP (1986) Reduction of the toxicity and mutagenicity of alkylating agents in mammalian cells harboring the Escherichia coli alkyltransferase gene. Proc Natl Acad Sci USA 83:6292–6296
Cooper AJ, Kelly SL (1987) DNA repair and mutagenesis in Saccharomyces cerevisiae. In: Wiseman A (ed) Theory and practice of mutagenicity testing in yeast. Ellis-Horwood, pp 73–114
Cox BS, Parry JM (1968) The isolation, genetics and survival characteristics of ultraviolet light-sensitive mutants in yeast. Mutat Res 6:37–55
Day RS, Ziolkowski CHJ, Scudiero DA, Meyer SA, Lubiniecki AS, Girardi AJ, Galloway SM, Bynum GD (1980) Defective repair of alkylated DNA by human tumour and SV40-transformed human cell strains. Nature 288:724–727
Foury F (1982) Endonucleases in yeast mitochondria: apurinic and managanese-stimulated deoxyribonuclease activities in the inner mitochondrial membrane of Saccharomyces cerevisiae. Eur J Biochem 124:253–259
Friedberg EC (1985) DNA repair. Freeman, New York
Game JC, Cox BS (1971) Allelism tests of mutants affecting sensitivity to radiation in yeast and a proposed nomenclature. Mutat Res 12:328–331
Game JC, Cox BS (1971) Epistatic interactions between four rad-loci in yeast. Mutat Res 16:353–362
Game JC, Cox BS (1973) Synergistic interactions between rad mutants in yeast. Mutat Res 20:35–44
Game JC, Mortimer RK (1974) A genetic study of X-ray sensitive mutants in yeast. Mutat Res 24:281–292
Haynes RH, Kunz BA (1981) DNA repair and mutagenesis in yeast. In: Strathern J, Jones E, Broach J (eds) The molecular biology of the yeast Saccharomyces Life cycle and inheritance. Cold Spring Harbor Laboratory, Cold Spring Harbor, pp 371–414
Hince TA, Neale S (1974) Effect of N-nitro-N-methylurea on viability and mutagenic response of repair deficient strains of E. coli. Mutat Res 22:235–242
Hoekstra MF, Malone RE (1986) Excision repair functions in Saccharomyces cerevisiae recognise and repair methylation of adenine by the Escherichia coli dam gene. Mol Cell Biol 6:3555–3558
Jeggo P, Defais M, Samson L, Schendel P (1978) The adaptive response of E. coli to low levels of alkylating agent: the role of polA in killing adaptation. Mol Gen Genet 162:299–305
Khan NA, Brendel M, Haynes RH (1970) Supersensitive double mutants in yeast. Mol Gen Genet 107:376–378
Kupeic M, Simchen G (1986) Regulation of the RAD6 gene of Saccharomyces cerevisiae in the mitotic cell cycle and in meiosis. Mol Gen Genet 203:538–543
Lawley PD (1974) Some chemical aspects of dose-response relationships in alkylation mutagenesis. Mutat Res 23:283–295
Lawley PD, Orr DJ, Jarman M (1975) Isolation and identification of products from alkylation of nucleic acids: ethyl- and isopropyl-purines. Biochem J 145:73–84
Lawrence CW (1982) Mutagenesis in Saccharomyces cerevisiae. Adv Genet 21:173–254
Lawrence CW, Christensen R (1979) Metabolic suppressors of trimethoprim and ultraviolet light sensitivites of Saccharomyces cerevisiae rad6 mutants. J Bacteriol 139:866–876
Lawrence CW, Stewart JW, Sherman F (1974) Specificity and frequency of UV-induced reversion of an iso-1-cytochrome c ochre mutant in radiation-sensitive strains of yeast. J Mol Biol 85:137–162
Lawrence CW, Stewart JW, Sherman F, Thomas FLX (1970) Mutagenesis in ultraviolet sensitive mutants of yeast. Genetics 64:s36-s37
Lindahl T (1982) DNA repair enzymes. Annu Rev Biochem 51:61–87
Lindahl T, Sedgwick B, Demple B, Karran P (1983) Enzymology and regulation of the adaptive response to alkylating agents. In: Friedberg EC, Bridges BA (eds) Cellular responses to DNA damage. Liss, New York
Maga JA, McEntree K (1985) Response of S. cerevisiae to N-methyl-N′-nitro-N-nitrosoguanidine: mutagenesis, survival and DDR gene expression. Mol Gen Genet 200:313–321
McCarthy JG, Edington BV, Schendel PF (1983) Inducible repair of phosphotriesters in Escherichia coli. Proc Natl Acad Sci USA 80:7380–7384
McCarthy TV, Karran P, Lindahl T (1984) Inducible repair of O-alkylated DNA pyrimidines in Escherichia coli. EMBO J 3:545–550
Miller RD, Prakash L, Prakash S (1982) Defective excision of pyrimidine dimers and interstrand DNA cross-links in rad7 and rad23 mutants of Saccharomyces cerevisiae. Mol Gen Genet 188:235–239
Nakabeppu Y, Sekiguchi M (1986) Regulatory mechanisms for induction of synthesis of repair enzymes in response to alkylating agents: Ada protein acts as a transcriptional regulator. Proc Natl Acad Sci USA 83:6297–6301
Nisson PE, Lawrence CW (1986) The isolation and characterisation of an alkylating agent sensitive yeast mutant, ngs1. Mutat Res 165:129–137
Prakash L (1974) Lack of chemically induced mutation in repair deficient mutants of yeast. Genetics 78:1101–1118
Prakash L (1977a) Repair of pyrimidine dimers in radiation-sensitive mutants rad3, rad4, rad6 and rad9 of S. cerevisiae. Mutat Res 45:13–20
Prakash L (1977b) Defective thymine dimer excision in radiation-sensitive mutants rad10 and rad16 of S. cerevisiae. Mol Gen Genet 152:125–128
Prakash L, Higgins D (1982) Role of DNA repair in ethylmethane-sulphonate-induced mutagenesis in S. cerevisiae. Carcinogenesis 3:439–444
Prakash L, Prakash S (1977) Isolation and characterisation of MMS-sensitive mutants of S. cerevisiae. Genetics 86:33–35
Prakash L, Prakash S (1979) Three additional genes involved in pyrimidine dimer removal in Saccharomyces cerevisiae: RAD7, RAD14 and MMS19. Mol Gen Genet 176:351–359
Resnick MA (1969) Genetic control of radiation sensitivity in S. cerevisiae. Genetics 62:519–531
Reynolds RJ, Friedberg EC (1981a) Molecular mechanisms of pyrimidine dimer excision in Saccharomyces cerevisiae. Incision of ultraviolet-irradiated doxyribonucleic acid in vivo. J Bacteriol 146:692–704
Reynolds RJ, Friedberg EC (1981b) Molecular mechanisms of pyrimidine dimer excision in Saccharomyces cerevisiae. Excision of dimers in cell extracts. J Bacteriol 147:705–708
Ruhland A, Brendel M (1979) Mutagenesis by cytostatic alkylating agents in yeast strains of differing repair capacities. Genetics 92:83–97
Samson L, Cairns J (1977) A new pathway for DNA repair in E. coli. Nature 267:281–283
Schendel PF, Edington BV, McCarthy JG, Todd ML (1983) Repair of alkylation damage in E. coli. In: Friedberg EC, Bridges BA (eds) Cellular responses to DNA damage. Liss, New York, pp 227–240
Sedgwick B, Lindahl T (1982) A common mechanisms for repair of O6 methylguanine and O6-ethylguanine in DNA. J Mol Biol 154:169–174
Shiloh Y, Kecker Y (1981) Kinetics of O6-methylguanine repair in human normal and ataxia telangiestasia cell lines and correlation of repair capacity with cellular sensitivity to methylating agents. Cancer Res 41:5114–5120
Singer B (1975) The chemical effects of nucleic acid alkylation and their relation to mutagenesis and carcinogenesis. Prog Nucl Acid Res Mol Biol 15:219–284
Singer B, Grunberger D (1983) Molecular biology of mutagens and carcinogens. Plenum Press, New York
Sklar RM, Strauss BS (1980) O6-Methylguanine removal by competent and incompetent human lymphoblastoid lines from the same male individual. Cancer Res 43:3316–3320
Sun L, Singer B (1975) The specificity of different classes of ethylating agents towards various sites of HeLa cell DNA in vitro and in vivo. Biochemistry 14:1795–1802
Szostak JM, Orr-Weaver TL, Rothstein RJ, Stahl FW (1983) The double-strand-break repair model for recombination. Cell 33:25–35
Teo I, Sedgwick B, Kilpatrick MW, McCarthy TV, Lindahl T (1986) The intracellular signal for induction of resistance of alkylating agents in E. coli. Cell 45:315–324
Tuite MF, Cox BS (1981) RAD6 + gene of Saccharomyces cerevisiae codes for 2 mutationally separable DNA repair functions. Mol Cell Biol 1:153–157
Unrau P, Wheatcroft R, Cox BS (1971) The excision of pyrimidine dimers from DNA of UV-irradiated yeast. Mol Gen Genet 113:359–362
Warren W, Lawley PD (1980) The removal of alkylation products from the DNA of E. coli cells treated with the carcinogens ENU and MNU: influence of growth conditions and DNA repair defects. Carcinogenesis 1:67–78
Waters R, Moustacchi E (1974) The disappearance of UV-induced pyrimidine dimers from the nuclear DNA of exponential and stationary phase cells of S. cerevisiae following post-irradiation treatments. Biochim Biophys Acta 353:407–419
Wilcox DR, Prakash L (1981) Incision and postincision steps of pyrimidine dimer removal in excision-defective mutants of S. cerevisiae. J Bacteriol 148:618–623
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Communicated by B.J. Kilbey
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Cooper, A.J., Waters, R. A complex pattern of sensitivity to simple monofunctional alkylating agents exists amongst the rad mutants of Saccharomyces cerevisiae . Mol Gen Genet 209, 142–148 (1987). https://doi.org/10.1007/BF00329849
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DOI: https://doi.org/10.1007/BF00329849