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Characterization of postreplication repair in Saccharomyces cerevisiae and effects of rad6, rad18, rev3 and rad52 mutations

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Summary

Postreplication repair of nuclear DNA was examined in an excision defective haploid strain of yeast lacking mitochondrial DNA (ral ρ0). The size of the DNA synthesized in cells exposed to various fluences of ultraviolet light (UV) corresponds approximately to the average interdimer distance in the parental DNA. Upon further incubation of cells following exposure to 2.5 J/m2, the DNA increases in size; by 4 h, it corresponds to DNA from uniformly labeled cells. The alkaline sucrose sedimentation pattern of DNA pulse labeled at various times after UV irradiation, for up to 4 h, does not change substantially, indicating that dimers continue to block DNA replication. A significant amount of postreplication repair requires de novo protein synthesis, as determined by its inhibition by cycloheximide. The rad6 mutant does not carry out postreplication repair, the rad18 and rad52 mutants show great inhibition while the rev3 mutation does not affect postreplication repair. Both recombinational and nonrecombinational repair mechanisms may function in postreplication repair and most of postreplication repair is error free.

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References

  • Boram WR, Roman H (1976) Recombination in Saccharomyces cerevisiae: a DNA repair mutation associated with elevated mitotic gene conversion. Proc Natl Acad Sci USA 73:2828–2832

    Google Scholar 

  • Buhl SN, Setlow RB, Regan JD (1972) Steps in DNA chain clongation and joining after ultra-violet irradiation of human cells. Int J Radiat Biol 22:417–424

    Google Scholar 

  • Buhl SN, Setlow RB, Regan JD (1973) Recovery of the ability to synthesize DNA in segments of normal size at long times after ultraviolet irradiation of human cells. Biophys J 13:1265–1275

    Google Scholar 

  • Buhl SN, Setlow RB, Regan JD (1974) DNA repair in Potorus tridactylus. Biophys J 14:791–803

    Google Scholar 

  • Burgi E, Hershey AD (1963) Sedimentation rate as a measure of molecular weights of DNA. Biophys J 3:309–321

    Google Scholar 

  • Clark RW, Wever GH, Wiberg JS (1980) High-molecular-weight DNA and the sedimentation coefficient: a new perspective based on DNA from T7 bacteriophage and two novel forms of T4 bacteriophage. J Virol 33:438–448

    Google Scholar 

  • Cox, BS, Game J (1974) Repair systems in Saccharomyces. Mutat Res 26:257–264

    Google Scholar 

  • Cox BS, Parry JM (1968) The isolation, genetics and survival characteristics of ultraviolet-light sensitive mutants in yeast. Mutat Res 6:37–55

    Google Scholar 

  • Cummins JE, Rusch HP (1966) Limited DNA synthesis in the absence of protein synthesis in Physarum polycephalum. J Cell Biol 31:577–583

    Google Scholar 

  • diCaprio L, Cox BS (1981) DNA synthesis in UV-irradiated yeast. Mutat Res 82:69–85

    Google Scholar 

  • Fabre F (1978) Induced intragenic recombination in yeast can occur during the G1 mitotic phase. Nature 272:795–798

    Google Scholar 

  • Game JC, Mortimer RK (1974) A genetic study of X-ray sensitive mutants in yeast. Mutat Res 24:281–292

    Google Scholar 

  • Game JC, Zamb TJ, Braun RJ, Resnick M, Roth RM (1980) The role of radiation (rad) genes in meiotic recombination in yeast. Genetics 94:51–68

    Google Scholar 

  • Goldring ES, Grossman LI, Krupnick D, Cryer DR, Marmur J (1970) The petite mutation in yeast. Loss of mitochondrial deoxyribonucleic acid during induction of petites with ethidium bromide. J Mol Biol 52:323–335

    Google Scholar 

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

    Google Scholar 

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

    Google Scholar 

  • Hill WE, Fangman WL (1973) Scission of Escherichia coli deoxyribonucleic acid in alkali. Biochem 12:1772–1774

    Google Scholar 

  • Hunnable EG, Cox BS (1971) The genetic control of dark recombination in yeast. Mutat Res 13:297–309

    Google Scholar 

  • Johnston LH, Williamson DH (1978) An alkaline sucrose gradient analysis of the mechanism of nuclear DNA synthesis in the yeast Saccharomyces cerevisiae. Mol Gen Genet 164:217–225

    Google Scholar 

  • Kutter EM, Wiberg JS (1968) Degradation of cytosine-containing bacterial and bacteriophage DNA after infection of Escherichia coli B with bacteriophage T4D wild-type and with mutants defective in genes 46, 47 and 56. J Mol Biol 38:395–411

    Google Scholar 

  • Lauer GD, Roberts TM, Klotz LC (1977) Determination of the nuclear DNA content of Saccharomyces cerevisiae and implications for the organization of DNA in yeast chromosomes. J Mol Biol 114:507–526

    Google Scholar 

  • Lawrence CW, Christensen RB (1976) UV mutagenesis in radiationsensitive strains of yeast. Genetics 82:207–232

    Google Scholar 

  • Lawrence CW, Christensen RB (1979) Ultraviolet-induced reversion of cyc1 alleles in radiation-sensitive strains of yeast. III. rev3 mutant strains. Genetics 92:397–408

    Google Scholar 

  • Lawrence CW, Stewart JW, Sherman F, Christensen R (1974) Specificity and frequency of ultraviolet-induced reversion of an iso-1-cytochrome c ochre mutant in radiation-sensitive strains of yeast. J Mol Biol 85:137–162

    Google Scholar 

  • Lehmann AR (1972) Postreplication repair of DNA in ultraviolet-irradiated mammalian cells. J Mol Biol, 66:319–337

    Google Scholar 

  • Lehmann AR, Kirk-Bell S (1972) Postreplication repair of DNA in ultraviolet-irradiated mammalian cells. No gaps in DNA synthesized late after ultraviolet irradiation. Eur J Biochem 31:438–445

    Google Scholar 

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

    Google Scholar 

  • Lemontt JF (1971b) Pathways of ultraviolet mutability in Saccharomyces cerevisiae. I. Some properties of double mutants involving uvs9 and rev. Mutat Res 13:311–317

    Google Scholar 

  • Lemontt JF (1971c) Pathways of ultraviolet mutability in Saccharomyces cerevisiae. II. The effect of rev genes on recombination. Mutat Res 13:319–326

    Google Scholar 

  • Lemontt JF (1973) Genes controlling ultraviolet mutability in yeast. Genetics Supplement 73:153–159

    Google Scholar 

  • Malone RE, Esposito RE (1980) The RAD52 gene is required for homothallic interconversion of mating types and spontaneous mitotic recombination in yeast. Proc Natl Acad Sci USA 77:503–507

    Google Scholar 

  • McKee RH, Lawrence CW (1979) Genetic analysis of gamma-ray mutagenesis in yeast. I. Reversion in radiation-sensitive strains. Genetics 93:361–373

    Google Scholar 

  • Meyn RE, Humphrey RM (1971) Deoxyribonucleic acid synthesis in ultraviolet-light irradiated Chinese hamster cells. Biophys J 11:295–301

    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 

  • Painter RB (1978) Does ultraviolet light enhance postreplication repair in mammalian cells. Nature 275:243–245

    Google Scholar 

  • Park SD, Cleaver JE (1979) Postreplication repair: questions of its definition and possible alteration in xeroderma pigmentosum cell strains. Proc Natl Acad Sci USA 76:3927–3931

    Google Scholar 

  • Prakash L (1974) Lack of chemically induced mutation in repairdeficient mutants of yeast. Genetics 78:1101–1118

    Google Scholar 

  • Prakash L (1975) Repair of pyrimidine dimers in nuclear and mitochondrial DNA of yeast irradiated with low doses of ultraviolet light. J Mol Biol 98:781–795

    Google Scholar 

  • Prakash L (1977) Repair of pyrimidine dimers in radiation-sensitive mutants rad3, rad4, rad6 and rad9 of Saccharomyces cerevisiae. Mutat Res 45:13–20

    Google Scholar 

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

    Google Scholar 

  • Prakash L, Taillon-Miller P (1981) Effects of the rad52 gene on sister chromatid recombination in Saccharomyces cerevisiae. Curr Genet 3:247–250

    Google Scholar 

  • Prakash S, Prakash L, Burke W, Montelone BA (1980) Effects of the RAD52 gene on recombination in Saccharomyces cerevisiae. Genetics 94:31–50

    Google Scholar 

  • Resnick MA (1969) Genetic control of radiation sensitivity in Saccharomyces cerevisiae. Genetics, 62:519–531

    Google Scholar 

  • Resnick MA (1975) The repair of double strand breaks in chromosomal DNA of yeast. In: Hanawalt PC and Setlow RB (eds) Molecular mechanisms for repair of DNA. Plenum Press, New York, p 549

    Google Scholar 

  • Reynolds RJ and 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 

  • Rivin CJ, Fangman WL (1980) Cell cycle phase expansion in nitrogenlimited cultures of Saccharomyces cerevisiae. J Cell Biol 85:96–107

    Google Scholar 

  • Rupp WD, Howard-Flanders P (1968) Discontinuities in the DNA synthesized in an excision-defective strain of Escherichia coli following ultraviolet irradiation. J Mol Biol 31:291–304

    Google Scholar 

  • Rupp WD, Wilde CE, Reno DL, Howard-Flanders P (1971) Exchanges between DNA strands in ultraviolet-irradiated Escherichia coli. J Mol Biol 61:25–44

    Google Scholar 

  • Rosenstein BS, Setlow RB (1980) DNA repair after ultraviolet irradiation of ICR2A frog cells. Pyrimidine dimers are long acting blocks to nascent DNA synthesis. Biophys J 31:196–206

    Google Scholar 

  • Sarasin AR, Hanawalt PC (1980) Replication of ultraviolet-irradiated Simian virus 40 in monkey kidney cells. J Mol Biol 138:299–319

    Google Scholar 

  • Sedgwick SG (1975) Inducible error-prone repair in Escherichia coli. Proc Natl Acad Sci USA 72:2753–2757

    Google Scholar 

  • Snow R (1967) Mutants of yeast sensitive to ultraviolet light. J Bacteriol 94:571–575

    Google Scholar 

  • Unrau P, Wheatcroft P, Cox B, Olive T (1973) The formation of pyrimidine dimers in the DNA of fungi and bacteria. Biochim Biophys Acta 312:626–632

    Google Scholar 

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

    Google Scholar 

Download references

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Authors and Affiliations

  1. Department of Radiation Biology and Biophysics, University of Rochester, School of Medicine and Dentistry, 14642, Rochester, New York, USA

    Louise Prakash

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  1. Louise Prakash
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Communicated by B.A. Bridges

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Prakash, L. Characterization of postreplication repair in Saccharomyces cerevisiae and effects of rad6, rad18, rev3 and rad52 mutations. Mol Gen Genet 184, 471–478 (1981). https://doi.org/10.1007/BF00352525

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  • DOI: https://doi.org/10.1007/BF00352525

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