Skip to main content
SpringerLink
Log in

Expression of the RAD1 and RAD3 genes of Saccharomyces cerevisiae is not affected by DNA damage or during the cell division cycle

  • Published:
Molecular and General Genetics MGG Aims and scope Submit manuscript

Summary

The RAD1 and RAD3 genes of Saccharomyces cerevisiae are required for excision repair of UV damaged DNA. In addition, the RAD3 gene is essential since rad3 deletions are recessive lethals. We have examined the induction of the RAD1 and RAD3 genes by DNA damage and during the cell division cycle. We have made fusions of the RAD1 and RAD3 genes with the Escherichia coli lacZ gene encoding β-galactosidase. β-galactosidase activity was measured in a Rad+ yeast strain containing the RAD1-lacZ or the RAD3-lacZ fusion, either in a multicopy replicating plasmid or as a single copy integrant resulting from transformation with an integrating plasmid which transforms yeast by homologous recombination in the yeast genome. No induction of β-galactosidase activity occurred after ultraviolet light (UV) or 4-nitroquinoline-1-oxide (NQO) treatment. Haploid cells of mating type a were synchronized by treatment with α factor and β-galactosidase activity was determined during different cell cycle stages. No change in β-galactosidase activity was observed in the strain containing the RAD1-lacZ or the RAD3-lacZ fusion integrated in the yeast genome.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Backendorf C, Brandsma JA, Kartasova T, van de Putte P (1983) In vivo regulation of the uvrA gene: role of the “-10” and “-35” promoter regions. Nucl Acids Res 11:5795–5810

    Google Scholar 

  • Botstein D, Falco SC, Stewart SE, Brennan M, Scherer S, Stinchcomb DT, Struhl K, Davis RW (1979) Sterile host yeasts (SHY): a eukaryotic system of biological containment for recombinant DNA experiments. Gene 8:17–24

    Google Scholar 

  • Casadaban MJ, Chou J, Cohen SN (1980) In vitro gene fusions that join an enzymatically active β-galactosidase segment to amino-terminal fragments of exogenous proteins: Escherichia coli plasmid vectors for the detection and cloning of translational initiation signals. J Bacteriol 143:971–980

    Google Scholar 

  • Easton AM, Kushner SR (1983) Transcription of the uvrD gene of Escherichia coli is controlled by the lexA repressor and by attenuation. Nucl Acids Res 11:8625–8640

    Google Scholar 

  • Guarente L (1983) Yeast promoters and lacZ fusions designed to study expression of cloned genes in yeast. In: Wu R, Grossman L, Moldave K, (eds) Methods in enzymology. Academic Press, New York, 101: pp 181–191

    Google Scholar 

  • 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 cerevisiae; life cycle and inheritance. Cold Spring Harbor, New York, pp 371–414

  • Higgins DR, Prakash S, Reynolds P, Prakash L (1983a) Molecular cloning and characterization of the RAD1 gene of Saccharomyces cerevisiae. Gene 26:119–126

    Google Scholar 

  • Higgins DR, Prakash S, Reynolds P, Polakowska R, Weber S, Prakash L (1983b) Isolation and characterization of the RAD3 gene of Saccharomyces cerevisiae and inviability of rad3 deletion mutants. Proc Natl Acad Sci USA 80:5680–5684

    Google Scholar 

  • Jachymczyk WJ, von Borstel RC, Mowat MRA, Hastings PJ (1981) Repair of interstrand cross-links in DNA of Saccharomyces cerevisiae requires two systems for DNA repair: the RAD3 system and the RAD51 system. Mol Gen Genet 182:196–205

    Google Scholar 

  • Johnston GC, Singer RA, Sharrow SO, Slater ML (1980) Cell division in the yeast Saccharomyces cerevisiae growing at different rates. Jour Gen Microbiol 118:479–484

    Google Scholar 

  • Kenyon CJ, Walker GC (1981) Expression of the E. coli uvrA gene is inducible. Nature 289:808–810

    Google Scholar 

  • Magaña-Schwencke N, Henriques JAP, Chanet R, Moustacchi E (1982) The fate of 8-methoxypsoralen photoinduced crosslinks in nuclear and mitochondrial yeast DNA: comparison of wild type and repair-deficient strains. Proc Natl Acad Sci USA 79:1722–1726

    Google Scholar 

  • McClanahan T, McEntee K (1984) Specific transcripts are elevated in Saccharomyces cerevisiae in response to DNA damage. Mol Cell Biol 4:2356–2363

    Google Scholar 

  • Miller RD, Prakash L, Prakash S (1982) Genetic control of excision of Saccharomyces cerevisiae interstrand DNA cross-links induced by psoralen plus near UV light. Mol Cell Biol 2:939–948

    Google Scholar 

  • Nakayama K, Irino N, Nakayama H (1983) recA + gene-dependent regulation of a uvrD::lacZ fusion in Escherichia coli K12. Mol Gen Genet 192:391–394

    Google Scholar 

  • Naumovski L, Friedberg EC (1983) A DNA repair gene required for the incision of damaged DNA is essential for viability in S. cerevisiae. Proc Natl Acad Sci USA 80:4818–4821

    Google Scholar 

  • Peterson TA, Prakash L, Prakash S, Osley MA, Reed SI (1985) Regulation of CDC9, the S. cerevisiae gene that encodes DNA ligase. Mol Cell Biol 5:226–235

    Google Scholar 

  • Reynolds RJ, Friedberg EC (1981) Molecular mechanisms of pyrimidine dimer excision in Saccharomyces cerevisiae: incision of ultraviolet-irradiated deoxyribonucleic acid in vivo. J Bacteriol 146:692–704

    Google Scholar 

  • Rose M, Botstein D (1983) Construction and use of gene fusions to lacZ (β-galactosidase) that are expressed in yeast. In: Wu R, Grossman L, Moldave K (eds) Methods in enzymology. Academic Press, New York, 101: pp 167–180

    Google Scholar 

  • Ruby SW, Szostak JW (1985) Specific Saccharomyces cerevisiae genes are expressed in response to DNA-damaging agents. Mol Cell Biol 5:75–84

    Google Scholar 

  • Sancar GB, Sancar A, Little JW, Rupp WD (1982) The uvrB gene of Escherichia coli has both lexA-repressed and lexA-independent promoters Cell 28:523–530

    Google Scholar 

  • Schendel PF, Fogliano M, Strausbaugh LD (1982) Regulation of the Escherichia coli K-12 uvrB operon. J Bacteriol 150:676–685

    Google Scholar 

  • Scherer S, Davis RW (1979) Replacement of chromosome segments with altered DNA sequence constructed in vitro. Proc Natl Acad Sci USA 76:4951–4955

    Google Scholar 

  • Siegel EC (1983) The Escherichia coli uvrD gene is inducible by DNA damage. Mol Gen Genet 191:397–400

    Google Scholar 

  • Williamson DH (1973) Replication of the nuclear genome in yeast does not require concommitant protein synthesis. Biochem Biophys Res Com 52:731–740

    Google Scholar 

  • Wilcox DR, Prakash L (1981) Incision and post incision steps of pyrimidine dimer removal in excision-defective mutants of Saccharomyces cerevisiae. J Bacteriol 148:618–623

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Biology, University of Rochester, 14627, Rochester, NY, USA

    Madan L. Nagpal, David R. Higgins & Satya Prakash

Authors
  1. Madan L. Nagpal
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. David R. Higgins
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Satya Prakash
    View author publications

    You can also search for this author in PubMed Google Scholar

Additional information

Communicated by C. Auerbach

Rights and permissions

Reprints and permissions

About this article

Cite this article

Nagpal, M.L., Higgins, D.R. & Prakash, S. Expression of the RAD1 and RAD3 genes of Saccharomyces cerevisiae is not affected by DNA damage or during the cell division cycle. Molec Gen Genet 199, 59–63 (1985). https://doi.org/10.1007/BF00327510

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00327510

Keywords

Search

Navigation