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. 1991 Dec 11;19(23):6427–6432. doi: 10.1093/nar/19.23.6427

Induction of S.cerevisiae MAG 3-methyladenine DNA glycosylase transcript levels in response to DNA damage.

J Chen 1, L Samson 1
PMCID: PMC329188  PMID: 1754379

Abstract

We previously showed that the expression of the Saccharomyces cerevisiae MAG 3-methyladenine (3MeA) DNA glycosylase gene, like that of the E. coli alkA 3MeA DNA glycosylase gene, is induced by alkylating agents. Here we show that the MAG induction mechanism differs from that of alkA, at least in part, because MAG mRNA levels are not only induced by alkylating agents but also by UV light and the UV-mimetic agent 4-nitroquinoline-1-oxide. Unlike some other yeast DNA-damage-inducible genes, MAG expression is not induced by heat shock. The S. cerevisiae MGT1 O6-methylguanine DNA methyltransferase is not involved in regulating MAG gene expression since MAG is efficiently induced in a methyltransferase deficient strain; similarly, MAG glycosylase deficient strains and four other methylmethane sulfonate sensitive strains were normal for alkylation-induced MAG gene expression. However, de novo protein synthesis is required to elevate MAG mRNA levels because MAG induction was abolished in the presence of cycloheximide. MAG mRNA levels were equally well induced in cycling and G1-arrested cells, suggesting that MAG induction is not simply due to a redistribution of cells into a part of the cell cycle which happens to express MAG at high levels, and that the inhibition of DNA synthesis does not act as the inducing signal.

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Selected References

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

  1. Berdal K. G., Bjørås M., Bjelland S., Seeberg E. Cloning and expression in Escherichia coli of a gene for an alkylbase DNA glycosylase from Saccharomyces cerevisiae; a homologue to the bacterial alkA gene. EMBO J. 1990 Dec;9(13):4563–4568. doi: 10.1002/j.1460-2075.1990.tb07909.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bücking-Throm E., Duntze W., Hartwell L. H., Manney T. R. Reversible arrest of haploid yeast cells in the initiation of DNA synthesis by a diffusible sex factor. Exp Cell Res. 1973 Jan;76(1):99–110. doi: 10.1016/0014-4827(73)90424-2. [DOI] [PubMed] [Google Scholar]
  3. Ceccoli J., Rosales N., Goldstein M., Yarosh D. B. Polyclonal antibodies against O6-methylguanine-DNA methyltransferase in adapted bacteria. Mutat Res. 1988 Nov;194(3):219–226. doi: 10.1016/0167-8817(88)90023-5. [DOI] [PubMed] [Google Scholar]
  4. Chen J., Derfler B., Maskati A., Samson L. Cloning a eukaryotic DNA glycosylase repair gene by the suppression of a DNA repair defect in Escherichia coli. Proc Natl Acad Sci U S A. 1989 Oct;86(20):7961–7965. doi: 10.1073/pnas.86.20.7961. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Chen J., Derfler B., Samson L. Saccharomyces cerevisiae 3-methyladenine DNA glycosylase has homology to the AlkA glycosylase of E. coli and is induced in response to DNA alkylation damage. EMBO J. 1990 Dec;9(13):4569–4575. doi: 10.1002/j.1460-2075.1990.tb07910.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Cole G. M., Schild D., Lovett S. T., Mortimer R. K. Regulation of RAD54- and RAD52-lacZ gene fusions in Saccharomyces cerevisiae in response to DNA damage. Mol Cell Biol. 1987 Mar;7(3):1078–1084. doi: 10.1128/mcb.7.3.1078. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Elledge S. J., Davis R. W. DNA damage induction of ribonucleotide reductase. Mol Cell Biol. 1989 Nov;9(11):4932–4940. doi: 10.1128/mcb.9.11.4932. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Elledge S. J., Davis R. W. Identification and isolation of the gene encoding the small subunit of ribonucleotide reductase from Saccharomyces cerevisiae: DNA damage-inducible gene required for mitotic viability. Mol Cell Biol. 1987 Aug;7(8):2783–2793. doi: 10.1128/mcb.7.8.2783. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. 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]
  10. Friedberg E. C. Deoxyribonucleic acid repair in the yeast Saccharomyces cerevisiae. Microbiol Rev. 1988 Mar;52(1):70–102. doi: 10.1128/mr.52.1.70-102.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hereford L. M., Osley M. A., Ludwig T. R., 2nd, McLaughlin C. S. Cell-cycle regulation of yeast histone mRNA. Cell. 1981 May;24(2):367–375. doi: 10.1016/0092-8674(81)90326-3. [DOI] [PubMed] [Google Scholar]
  12. Hooley P., Shawcross S. G., Strike P. An adaptive response to alkylating agents in Aspergillus nidulans. Curr Genet. 1988 Nov;14(5):445–449. doi: 10.1007/BF00521267. [DOI] [PubMed] [Google Scholar]
  13. Hurd H. K., Roberts J. W. Upstream regulatory sequences of the yeast RNR2 gene include a repression sequence and an activation site that binds the RAP1 protein. Mol Cell Biol. 1989 Dec;9(12):5359–5372. doi: 10.1128/mcb.9.12.5359. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Johnston L. H., White J. H., Johnson A. L., Lucchini G., Plevani P. The yeast DNA polymerase I transcript is regulated in both the mitotic cell cycle and in meiosis and is also induced after DNA damage. Nucleic Acids Res. 1987 Jul 10;15(13):5017–5030. doi: 10.1093/nar/15.13.5017. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Jones J. S., Prakash L., Prakash S. Regulated expression of the Saccharomyces cerevisiae DNA repair gene RAD7 in response to DNA damage and during sporulation. Nucleic Acids Res. 1990 Jun 11;18(11):3281–3285. doi: 10.1093/nar/18.11.3281. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Jones J. S., Prakash L. Transcript levels of the Saccharomyces cerevisiae DNA repair gene RAD18 increase in UV irradiated cells and during meiosis but not during the mitotic cell cycle. Nucleic Acids Res. 1991 Feb 25;19(4):893–898. doi: 10.1093/nar/19.4.893. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. 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]
  18. Laval F. Induction of proteins involved in the repair of alkylated bases in mammalian cells by DNA-damaging agents. Mutat Res. 1990 Nov-Dec;233(1-2):211–218. doi: 10.1016/0027-5107(90)90164-y. [DOI] [PubMed] [Google Scholar]
  19. 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]
  20. Madura K., Prakash S. Transcript levels of the Saccharomyes cerevisiae DNA repair gene RAD23 increase in response to UV light and in meiosis but remain constant in the mitotic cell cycle. Nucleic Acids Res. 1990 Aug 25;18(16):4737–4742. doi: 10.1093/nar/18.16.4737. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. 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]
  22. McClanahan T., McEntee K. DNA damage and heat shock dually regulate genes in Saccharomyces cerevisiae. Mol Cell Biol. 1986 Jan;6(1):90–96. doi: 10.1128/mcb.6.1.90. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. McClanahan T., McEntee K. Specific transcripts are elevated in Saccharomyces cerevisiae in response to DNA damage. Mol Cell Biol. 1984 Nov;4(11):2356–2363. doi: 10.1128/mcb.4.11.2356. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Nakabeppu Y., Miyata T., Kondo H., Iwanaga S., Sekiguchi M. Structure and expression of the alkA gene of Escherichia coli involved in adaptive response to alkylating agents. J Biol Chem. 1984 Nov 25;259(22):13730–13736. [PubMed] [Google Scholar]
  25. Peterson T. A., Prakash L., Prakash S., Osley M. A., Reed S. I. Regulation of CDC9, the Saccharomyces cerevisiae gene that encodes DNA ligase. Mol Cell Biol. 1985 Jan;5(1):226–235. doi: 10.1128/mcb.5.1.226. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. 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]
  27. Prakash L., Prakash S. Isolation and characterization of MMS-sensitive mutants of Saccharomyces cerevisiae. Genetics. 1977 May;86(1):33–55. doi: 10.1093/genetics/86.1.33. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Robinson G. W., Nicolet C. M., Kalainov D., Friedberg E. C. A yeast excision-repair gene is inducible by DNA damaging agents. Proc Natl Acad Sci U S A. 1986 Mar;83(6):1842–1846. doi: 10.1073/pnas.83.6.1842. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Ruby S. W., Szostak J. W., Murray A. W. Cloning regulated yeast genes from a pool of lacZ fusions. Methods Enzymol. 1983;101:253–269. doi: 10.1016/0076-6879(83)01019-8. [DOI] [PubMed] [Google Scholar]
  30. Ruby S. W., Szostak J. W. Specific Saccharomyces cerevisiae genes are expressed in response to DNA-damaging agents. Mol Cell Biol. 1985 Jan;5(1):75–84. doi: 10.1128/mcb.5.1.75. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Samson L., Cairns J. A new pathway for DNA repair in Escherichia coli. Nature. 1977 May 19;267(5608):281–283. doi: 10.1038/267281a0. [DOI] [PubMed] [Google Scholar]
  32. Sassanfar M., Dosanjh M. K., Essigmann J. M., Samson L. Relative efficiencies of the bacterial, yeast, and human DNA methyltransferases for the repair of O6-methylguanine and O4-methylthymine. Suggestive evidence for O4-methylthymine repair by eukaryotic methyltransferases. J Biol Chem. 1991 Feb 15;266(5):2767–2771. [PubMed] [Google Scholar]
  33. Sassanfar M., Samson L. Identification and preliminary characterization of an O6-methylguanine DNA repair methyltransferase in the yeast Saccharomyces cerevisiae. J Biol Chem. 1990 Jan 5;265(1):20–25. [PubMed] [Google Scholar]
  34. Sebastian J., Kraus B., Sancar G. B. Expression of the yeast PHR1 gene is induced by DNA-damaging agents. Mol Cell Biol. 1990 Sep;10(9):4630–4637. doi: 10.1128/mcb.10.9.4630. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Siede W., Robinson G. W., Kalainov D., Malley T., Friedberg E. C. Regulation of the RAD2 gene of Saccharomyces cerevisiae. Mol Microbiol. 1989 Dec;3(12):1697–1707. doi: 10.1111/j.1365-2958.1989.tb00155.x. [DOI] [PubMed] [Google Scholar]
  36. Storz G., Tartaglia L. A., Ames B. N. Transcriptional regulator of oxidative stress-inducible genes: direct activation by oxidation. Science. 1990 Apr 13;248(4952):189–194. doi: 10.1126/science.2183352. [DOI] [PubMed] [Google Scholar]
  37. Storz G., Tartaglia L. A., Farr S. B., Ames B. N. Bacterial defenses against oxidative stress. Trends Genet. 1990 Nov;6(11):363–368. doi: 10.1016/0168-9525(90)90278-e. [DOI] [PubMed] [Google Scholar]
  38. Struhl K. Molecular mechanisms of transcriptional regulation in yeast. Annu Rev Biochem. 1989;58:1051–1077. doi: 10.1146/annurev.bi.58.070189.005155. [DOI] [PubMed] [Google Scholar]
  39. Teo I., Sedgwick B., Kilpatrick M. W., McCarthy T. V., Lindahl T. The intracellular signal for induction of resistance to alkylating agents in E. coli. Cell. 1986 Apr 25;45(2):315–324. doi: 10.1016/0092-8674(86)90396-x. [DOI] [PubMed] [Google Scholar]
  40. Treger J. M., Heichman K. A., McEntee K. Expression of the yeast UB14 gene increases in response to DNA-damaging agents and in meiosis. Mol Cell Biol. 1988 Mar;8(3):1132–1136. doi: 10.1128/mcb.8.3.1132. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Vaughan P., Sedgwick B., Hall J., Gannon J., Lindahl T. Environmental mutagens that induce the adaptive response to alkylating agents in Escherichia coli. Carcinogenesis. 1991 Feb;12(2):263–268. doi: 10.1093/carcin/12.2.263. [DOI] [PubMed] [Google Scholar]
  42. Volkert M. R., Nguyen D. C., Beard K. C. Escherichia coli gene induction by alkylation treatment. Genetics. 1986 Jan;112(1):11–26. doi: 10.1093/genetics/112.1.11. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Volkert M. R., Nguyen D. C. Induction of specific Escherichia coli genes by sublethal treatments with alkylating agents. Proc Natl Acad Sci U S A. 1984 Jul;81(13):4110–4114. doi: 10.1073/pnas.81.13.4110. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Walker G. C. Mutagenesis and inducible responses to deoxyribonucleic acid damage in Escherichia coli. Microbiol Rev. 1984 Mar;48(1):60–93. doi: 10.1128/mr.48.1.60-93.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Xiao W., Derfler B., Chen J., Samson L. Primary sequence and biological functions of a Saccharomyces cerevisiae O6-methylguanine/O4-methylthymine DNA repair methyltransferase gene. EMBO J. 1991 Aug;10(8):2179–2186. doi: 10.1002/j.1460-2075.1991.tb07753.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Yagle K., McEntee K. The DNA damage-inducible gene DIN1 of Saccharomyces cerevisiae encodes a regulatory subunit of ribonucleotide reductase and is identical to RNR3. Mol Cell Biol. 1990 Oct;10(10):5553–5557. doi: 10.1128/mcb.10.10.5553. [DOI] [PMC free article] [PubMed] [Google Scholar]

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