Skip to main content
Log in

Analysis of mutagenic DNA repair in a thermoconditional repair mutant of Saccharomyces cerevisiae

I. Influence of cycloheximide on UV-irradiated stationary phase rev2 ts cells

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

Summary

Using the thermoconditional yeast mutant rev2 ts that controls an apparently site-specific step of mutagenic DNA repair it was possible to measure the time course of REV2 dependent UV-induced reversion of the ochre allele his5-2 and recovery of survival for UV-treated stationary phase cells: due to the rev2 ts coded protein being active at 23° C, survival and mutation frequencies increased with duration of incubation under permissive conditions in growth medium before the temperature was shifted to 36° C (restrictive temperature). This increase was abolished in the presence of the protein synthesis inhibitor, cycloheximide. Furthermore, the REV2 dependent recovery of survival could be blocked or nearly blocked by cycloheximide added at any time during repair. Therefore, REV2 dependent repair can be characterized as a process requiring concomitant protein synthesis. These findings give further support to the concept that in yeast, mutagenesis involves UV inducible components of DNA repair.

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

Access this article

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

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Budd M, Mortimer RK (1982) Repair of double-strand breaks in a temperature conditional radiation-sensitive mutant of Saccharomyces cerevisiae. Mutat Res 103:19–24

    Google Scholar 

  • Cooper PK, Hunt JG (1978) Alternative pathways for excision and resynthesis in Escherichia coli: DNA polymerase III role? In: Hanawalt PC, Friedberg EC, Fox CF (eds) ICN-UCLA symposia on molecular and cellular biology. Academic Press, New York, pp 255–260

    Google Scholar 

  • Defais M, Calliet-Fauquet P, Fox MS, Radman M (1976) Induction kinetics of mutagenic DNA repair activity in E. coli following ultraviolet irradiation. Mol Gen Genet 148:125–130

    Google Scholar 

  • Eckardt F, Moustacchi E, Haynes RH (1978) On the inducibility or error-prone repair in yeast. In: Hanawalt PC, Friedberg EC, Fox CF (eds) ICN-UCLA symposia on molecular and cellular biology. Academic Press, New York, pp 421–423

    Google Scholar 

  • Eckardt F, Teh SJ, Haynes RH (1980) Heteroduplex repair as an intermediate step of UV mutagenesis in yeast. Genetics 95:63–80

    Google Scholar 

  • Fabre F (1982) Vegetative transmission of the induced recombination ability in yeast. In: Abstracts of the XI. conference on yeast genetics and molecular biology, Montpellier, p 172

  • Fabre F, Roman H (1977) Genetic evidence for inducibility of recombination competence in yeast. Proc Natl Acad Sci USA 74:1667–1671

    Google Scholar 

  • Fäth WW, Brendel M (1974) Specific DNA-labelling by exogenous thymidine-5′-monophosphate in Saccharomyces cerevisiae. Mol Gen Genet 131:57–67

    Google Scholar 

  • Game JC, Cox BS (1971) Allelism tests of mutants affecting sensitivity to radiation in yeast and a proposed nomenclature. Mutat Res 12:328–331

    Google Scholar 

  • Hanawalt PC, Cooper PK, Ganesan AK, Smith CA (1979) DNA repair in bacteria and mammalian cells. Annu Rev Biochem 48:783–836

    Google Scholar 

  • Hawthorne DC, Leupold U (1974) Suppressor mutations in yeast. Curr Top Microbiol Immunol 64:1–47

    Google Scholar 

  • Haynes RH, Eckardt F (1979) Complexity of DNA repair in a simple eucaryote. In: Okada S, Imamura M, Terasima T, Yamaguchi H (eds) Radiation research, Proc. 6th Int. Congress of radiation research. Toppan Printing Co, Tokyo, pp 454–461

    Google Scholar 

  • Haynes RH, Kunz BA (1981) DNA repair and mutagenesis in yeast. In: Strathern JN, Jones EW, Broach JR (eds) The molecular biology of the yeast Saccharomyces. Cold Spring Harbor Laboratory Publications, New York, pp 371–414

    Google Scholar 

  • Hereford LM, Hartwell LH (1973) Role of protein synthesis in the replication of yeast DNA. Nature New Biol 244:129–131

    Google Scholar 

  • Ho KSY, Mortimer RK (1975) Two mutations which confer temperature-sensitive radiation sensitivity in the yeast Saccharomyces cerevisiae. Mutat Res 33:157–164

    Google Scholar 

  • James AP, Kilbey BJ (1977) The timing of UV mutagenesis in yeast: a pedigree analysis of induced recessive mutation. Genetics 87:237–248

    Google Scholar 

  • Lawrence CW (1982) Mechanisms of induced mutagenesis in yeast. In: Sugimura T, Kondo S, Takebe H (eds) Environmental mutagens and carcinogens. Alan R Liss Inc, Tokyo, pp 128–136

    Google Scholar 

  • lawrence CW, Christensen RB (1978) Ultraviolet-induced reversion of cycl alleles in radiation sensitive strains of yeast. II. rev2 mutant strains. Genetics 90:213–226

    Google Scholar 

  • Lawrence CW, Christensen RB (1982) The mechanism of untargeted mutagenesis in UV-irradiated yeast. Mol Gen Genet 186:1–9

    Google Scholar 

  • Lee MG, Yarranton GT (1982) Inducible DNA repair in Ustilago maydis. Mol Gen Genet 185:245–250

    Google Scholar 

  • Lemontt JF (1971) Mutants of yeast defective in mutation induced by ultraviolet light. Genetics 68:21–33

    Google Scholar 

  • Lemontt JF (1972) Induction of forward mutations in mutationally defective yeast. Mol Gen Genet 119:27–42

    Google Scholar 

  • lemontt JF (1980) Genetic and physiological factors affecting repair and mutagenesis in yeast. In: Generoso WM, Shelby MD, de Serres FJ (eds) DNA repair and mutagenesis in Eukaryotes. Plenum Press, New York, pp 85–120

    Google Scholar 

  • Little JW, Edmiston SH, Pacelli LZ, Mount DW (1980) Cleavage of the Escherichia coli lexA protein by the recA protease. Proc Natl Acad Sci USA 77:3223–3229

    Google Scholar 

  • McKee RH, Lawrence CW (1979) Genetic analysis of gamma ray mutagenesis in yeast. II. Allele specific control of mutagenesis. Genetics 93:375–381

    Google Scholar 

  • Montelone BA, Prakash S, Prakash L (1981) Recombination and mutagenesis in rad6 mutants of Saccharomyces cerevisiae: evidence for multiple functions of the RAD6 gene. Mol Gen Genet 184:410–415

    Google Scholar 

  • Mortimer RK, Hawthorne DC (1969) Yeast genetics. In: Rose AH, Harrison JS (eds) The yeasts, vol I. Academic Press, London New York, pp 385–460

    Google Scholar 

  • Nasim A, Hannan MA, Nestmann ER (1981) Pure and mosaic clones — a reflection of differences in mechanisms of mutagenesis by different agents in Saccharomyces cerevisiae. Can J Genet Cytol 23:79–79

    Google Scholar 

  • Njagi GDE, Kilbey BJ (1982) cdc7-1 a temperature sensitive cellcycle mutant which interferes with induced mutagenesis in Saccharomyces cerevisiae. Mol Gen Genet 186:478–481

    Google Scholar 

  • Prakash L (1981) Characterization of postreplication repair in Saccharomyces cerevisiae and effects of rad6, rad18, rev3 and rad52 mutations. Mol Gen Genet 184:471–478

    Google Scholar 

  • Prakash L, Prakash S (1980) Genetic analysis of error-prone repair systems in Saccharomyces cerevisiae. In: Generoso WM, Shelby MD, de Serres FJ (eds) DNA repair and mutagenesis in Eukaryotes. Plenum Press, New York, pp 141–158

    Google Scholar 

  • Radman M (1980) Is there SOS induction in mammalian cells? Photochem Photobiol 32:823–830

    Google Scholar 

  • Schendel PF (1981) Inducible repair systems and their implications for toxicology. CRC Crit Rev Toxicol 8:311–362

    Google Scholar 

  • Schwencke J, Moustacchi E (1982) Proteolytic activities in yeast after UV irradiation. I. Variation in proteinase levels in repair proficient RAD + strains. Mol Gen Genet 185:290–295

    Google Scholar 

  • Siede W, Brendel M (1981) Isolation and characterization of yeast mutants with thermoconditional sensitivity to the bifunctional alkylating agent nitrogen mustard. Curr Genet 4:145–149

    Google Scholar 

  • Siede W, Brendel M (1982) Mutant gene snm2-1 ts, conferring thermoconditional mutagen sensitivity in Saccharomyces cerevisiae, is allelic with RAD5. Curr Genet 5:93–95

    Google Scholar 

  • Siede W, Eckardt F, Brendel M (1983) Analysis of mutagenic DNA repair in a thermoconditional repair mutant of Saccharomyces cerevisiae. II. Influence of cycloheximide on UV irradiated exponentially growing rev2 ts cells. Mol Gen Genet 190:416

    Google Scholar 

  • Tuite MF, Cox BS (1981) RAD6 + gene of Saccharomyces cerevisiae codes for two mutationally separable deoxyribonucleic acid repair functions. Mol Cell Biol 1:153–157

    Google Scholar 

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

    Google Scholar 

  • Witkin EM (1974) Thermal enhancement of ultraviolet mutability in a tif-1 uvr A derivative of Escherichia coli B/r: Evidence that ultraviolet mutagenesis depends upon an inducible function. Proc Natl Acad Sci USA 71:1930–1934

    Google Scholar 

  • Witkin EM (1976) Ultraviolet mutagenesis and inducible DNA repair in Escherichia coli. Bacteriol Rev 40:869–907

    Google Scholar 

  • Witkin EM, George DL (1973) Ultraviolet mutagenesis and inductible DNA repair in polA and uvrA polA derivatives of Escherichia coli B/r: Evidence for an inducible error-prone repair system. Genetics (suppl) 73:91–108

    Google Scholar 

  • Yarranton GT, Lee MG (1982) UV inducible responses in simple eukaryotes. In: Abstracts of the XI. conference on yeast genetics and molecular biology, Montpellier, p 25

Download references

Author information

Authors and Affiliations

Authors

Additional information

Communicated by F. Kaudewitz

Rights and permissions

Reprints and permissions

About this article

Cite this article

Siede, W., Eckardt, F. & Brendel, M. Analysis of mutagenic DNA repair in a thermoconditional repair mutant of Saccharomyces cerevisiae . Mol Gen Genet 190, 406–412 (1983). https://doi.org/10.1007/BF00331068

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00331068

Keywords

Navigation