Skip to main content
NCBI home page
Molecular and Cellular Biology logoLink to Molecular and Cellular Biology
. 1985 Jan;5(1):75–84. doi: 10.1128/mcb.5.1.75

Specific Saccharomyces cerevisiae genes are expressed in response to DNA-damaging agents.

S W Ruby, J W Szostak
PMCID: PMC366680  PMID: 3920512

Abstract

When exposed to DNA-damaging agents, the yeast Saccharomyces cerevisiae induces the expression of at least six specific genes. We have previously identified one damage inducible (DIN) gene as a gene fusion (din-lacZ fusion) whose expression increases in response to DNA-damaging treatments. We describe here the identification of five additional DIN genes as din-lacZ fusions and the responses of all six DIN genes to DNA-damaging agents. Northern blot analyses of the transcripts of two of the DIN genes show that their levels increase after exposure to DNA-damaging agents. Five of the din-lacZ fusions are induced in S. cerevisiae cells exposed to UV light, gamma rays, methotrexate, or alkylating agents. One of the din-lacZ fusions is induced by either UV or methotrexate but not by the other agents. This finding suggests that there are sets of DIN genes that are regulated differently.

Full text

PDF
75

Images in this article

81Image
on p.81
82Image
on p.82

Selected References

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

  1. Alwine J. C., Kemp D. J., Parker B. A., Reiser J., Renart J., Stark G. R., Wahl G. M. Detection of specific RNAs or specific fragments of DNA by fractionation in gels and transfer to diazobenzyloxymethyl paper. Methods Enzymol. 1979;68:220–242. doi: 10.1016/0076-6879(79)68017-5. [DOI] [PubMed] [Google Scholar]
  2. Bagg A., Kenyon C. J., Walker G. C. Inducibility of a gene product required for UV and chemical mutagenesis in Escherichia coli. Proc Natl Acad Sci U S A. 1981 Sep;78(9):5749–5753. doi: 10.1073/pnas.78.9.5749. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Barker D. G., Johnston L. H. Saccharomyces cerevisiae cdc9, a structural gene for yeast DNA ligase which complements Schizosaccharomyces pombe cdc17. Eur J Biochem. 1983 Aug 1;134(2):315–319. doi: 10.1111/j.1432-1033.1983.tb07568.x. [DOI] [PubMed] [Google Scholar]
  4. Brunborg G., Resnick M. A., Williamson D. H. Cell-cycle-specific repair of DNA double strand breaks in Saccharomyces cerevisiae. Radiat Res. 1980 Jun;82(3):547–558. [PubMed] [Google Scholar]
  5. Cairns J., Robins P., Sedgwick B., Talmud P. The inducible repair of alkylated DNA. Prog Nucleic Acid Res Mol Biol. 1981;26:237–244. doi: 10.1016/s0079-6603(08)60408-0. [DOI] [PubMed] [Google Scholar]
  6. Casadaban M. J., Chou J., Cohen S. N. In vitro gene fusions that join an enzymatically active beta-galactosidase segment to amino-terminal fragments of exogenous proteins: Escherichia coli plasmid vectors for the detection and cloning of translational initiation signals. J Bacteriol. 1980 Aug;143(2):971–980. doi: 10.1128/jb.143.2.971-980.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Casadaban M. J., Cohen S. N. Lactose genes fused to exogenous promoters in one step using a Mu-lac bacteriophage: in vivo probe for transcriptional control sequences. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4530–4533. doi: 10.1073/pnas.76.9.4530. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Clewell D. B., Helinski D. R. Supercoiled circular DNA-protein complex in Escherichia coli: purification and induced conversion to an opern circular DNA form. Proc Natl Acad Sci U S A. 1969 Apr;62(4):1159–1166. doi: 10.1073/pnas.62.4.1159. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Crabeel M., Huygen R., Cunin R., Glansdorff N. The promoter region of the arg3 gene in Saccharomyces cerevisiae: nucleotide sequence and regulation in an arg3-lacZ gene fusion. EMBO J. 1983;2(2):205–212. doi: 10.1002/j.1460-2075.1983.tb01406.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Demple B., Halbrook J. Inducible repair of oxidative DNA damage in Escherichia coli. Nature. 1983 Aug 4;304(5925):466–468. doi: 10.1038/304466a0. [DOI] [PubMed] [Google Scholar]
  11. Fabre F., Roman H. Genetic evidence for inducibility of recombination competence in yeast. Proc Natl Acad Sci U S A. 1977 Apr;74(4):1667–1671. doi: 10.1073/pnas.74.4.1667. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Guarente L., Ptashne M. Fusion of Escherichia coli lacZ to the cytochrome c gene of Saccharomyces cerevisiae. Proc Natl Acad Sci U S A. 1981 Apr;78(4):2199–2203. doi: 10.1073/pnas.78.4.2199. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. 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]
  14. Higgins D. R., Prakash S., Reynolds P., Polakowska R., Weber S., Prakash L. Isolation and characterization of the RAD3 gene of Saccharomyces cerevisiae and inviability of rad3 deletion mutants. Proc Natl Acad Sci U S A. 1983 Sep;80(18):5680–5684. doi: 10.1073/pnas.80.18.5680. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Higgins D. R., Prakash S., Reynolds P., Prakash L. Molecular cloning and characterization of the RAD1 gene of Saccharomyces cerevisiae. Gene. 1983 Dec;26(2-3):119–126. doi: 10.1016/0378-1119(83)90181-6. [DOI] [PubMed] [Google Scholar]
  16. Kenyon C. J., Walker G. C. DNA-damaging agents stimulate gene expression at specific loci in Escherichia coli. Proc Natl Acad Sci U S A. 1980 May;77(5):2819–2823. doi: 10.1073/pnas.77.5.2819. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Kunz B. A., Barclay B. J., Game J. C., Little J. G., Haynes R. H. Induction of mitotic recombination in yeast by starvation for thymine nucleotides. Proc Natl Acad Sci U S A. 1980 Oct;77(10):6057–6061. doi: 10.1073/pnas.77.10.6057. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Kupiec M., Simchen G. Cloning and mapping of the RAD50 gene of Saccharomyces cerevisiae. Mol Gen Genet. 1984;193(3):525–531. doi: 10.1007/BF00382094. [DOI] [PubMed] [Google Scholar]
  19. Lawrence C. W. Mutagenesis in Saccharomyces cerevisiae. Adv Genet. 1982;21:173–254. doi: 10.1016/s0065-2660(08)60299-0. [DOI] [PubMed] [Google Scholar]
  20. Leaper S., Resnick M. A., Holliday R. Repair of double-strand breaks and lethal damage in DNA of Ustilago maydis. Genet Res. 1980 Jun;35(3):291–307. doi: 10.1017/s0016672300014154. [DOI] [PubMed] [Google Scholar]
  21. Lehrach H., Diamond D., Wozney J. M., Boedtker H. RNA molecular weight determinations by gel electrophoresis under denaturing conditions, a critical reexamination. Biochemistry. 1977 Oct 18;16(21):4743–4751. doi: 10.1021/bi00640a033. [DOI] [PubMed] [Google Scholar]
  22. Lemontt J. F. Genetic and physiological factors affecting repair and mutagenesis in yeast. Basic Life Sci. 1980;15:85–120. doi: 10.1007/978-1-4684-3842-0_7. [DOI] [PubMed] [Google Scholar]
  23. Lindahl T. DNA repair enzymes. Annu Rev Biochem. 1982;51:61–87. doi: 10.1146/annurev.bi.51.070182.000425. [DOI] [PubMed] [Google Scholar]
  24. Little J. W., Mount D. W. The SOS regulatory system of Escherichia coli. Cell. 1982 May;29(1):11–22. doi: 10.1016/0092-8674(82)90085-x. [DOI] [PubMed] [Google Scholar]
  25. Martinez-Arias A. E., Casadaban M. J. Fusion of the Saccharomyces cerevisiae leu2 gene to an Escherichia coli beta-galactosidase gene. Mol Cell Biol. 1983 Apr;3(4):580–586. doi: 10.1128/mcb.3.4.580. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Naumovski L., Friedberg E. C. Molecular cloning of eucaryotic genes required for excision repair of UV-irradiated DNA: isolation and partial characterization of the RAD3 gene of Saccharomyces cerevisiae. J Bacteriol. 1982 Oct;152(1):323–331. doi: 10.1128/jb.152.1.323-331.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Naumovski L., Friedberg E. C. Saccharomyces cerevisiae RAD2 gene: isolation, subcloning, and partial characterization. Mol Cell Biol. 1984 Feb;4(2):290–295. doi: 10.1128/mcb.4.2.290. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Orr-Weaver T. L., Szostak J. W., Rothstein R. J. Genetic applications of yeast transformation with linear and gapped plasmids. Methods Enzymol. 1983;101:228–245. doi: 10.1016/0076-6879(83)01017-4. [DOI] [PubMed] [Google Scholar]
  29. Orr-Weaver T. L., Szostak J. W., Rothstein R. J. Yeast transformation: a model system for the study of recombination. Proc Natl Acad Sci U S A. 1981 Oct;78(10):6354–6358. doi: 10.1073/pnas.78.10.6354. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Osley M. A., Hereford L. Identification of a sequence responsible for periodic synthesis of yeast histone 2A mRNA. Proc Natl Acad Sci U S A. 1982 Dec;79(24):7689–7693. doi: 10.1073/pnas.79.24.7689. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Prakash L. The relation between repair of DNA and radiation and chemical mutagenesis in Saccharomyces cerevisiae. Mutat Res. 1976 Dec;41(2-3):241–248. doi: 10.1016/0027-5107(76)90097-x. [DOI] [PubMed] [Google Scholar]
  32. Quillardet P., Huisman O., D'Ari R., Hofnung M. SOS chromotest, a direct assay of induction of an SOS function in Escherichia coli K-12 to measure genotoxicity. Proc Natl Acad Sci U S A. 1982 Oct;79(19):5971–5975. doi: 10.1073/pnas.79.19.5971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Rigby P. W., Dieckmann M., Rhodes C., Berg P. Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol. 1977 Jun 15;113(1):237–251. doi: 10.1016/0022-2836(77)90052-3. [DOI] [PubMed] [Google Scholar]
  34. Rose M., Casadaban M. J., Botstein D. Yeast genes fused to beta-galactosidase in Escherichia coli can be expressed normally in yeast. Proc Natl Acad Sci U S A. 1981 Apr;78(4):2460–2464. doi: 10.1073/pnas.78.4.2460. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. 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]
  36. 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]
  37. Siede W., Eckardt F., Brendel M. Analysis of mutagenic DNA repair in a thermoconditional repair mutant of Saccharomyces cerevisiae. I. Influence of cycloheximide on UV-irradiated stationary phase rev2ts cells. Mol Gen Genet. 1983;190(3):406–412. doi: 10.1007/BF00331068. [DOI] [PubMed] [Google Scholar]
  38. Silverman S. J., Rose M., Botstein D., Fink G. R. Regulation of HIS4-lacZ fusions in Saccharomyces cerevisiae. Mol Cell Biol. 1982 Oct;2(10):1212–1219. doi: 10.1128/mcb.2.10.1212. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  40. Stadler D., Moyer R. Induced repair of genetic damage in Neurospora. Genetics. 1981 Aug;98(4):763–774. doi: 10.1093/genetics/98.4.763. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Struhl K., Davis R. W. A physical, genetic and transcriptional map of the cloned his3 gene region of Saccharomyces cerevisiae. J Mol Biol. 1980 Jan 25;136(3):309–332. doi: 10.1016/0022-2836(80)90376-9. [DOI] [PubMed] [Google Scholar]
  42. Struhl K., Davis R. W. Transcription of the his3 gene region in Saccharomyces cerevisiae. J Mol Biol. 1981 Nov 5;152(3):535–552. doi: 10.1016/0022-2836(81)90267-9. [DOI] [PubMed] [Google Scholar]
  43. 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]
  44. Witkin E. M. Ultraviolet mutagenesis and inducible DNA repair in Escherichia coli. Bacteriol Rev. 1976 Dec;40(4):869–907. doi: 10.1128/br.40.4.869-907.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Molecular and Cellular Biology are provided here courtesy of Taylor & Francis

RESOURCES