Summary
We have investigated the effects of UV irradiation of Saccharomyces cerevisiae in order to distinguish whether UV-induced recombination results from the induction of enzymes required for homologous recombination, of the production of substrate sites for recombination containing regions of DNA damage. We utilized split-dose experiments to investigate the induction of proteins required for survival, gene conversion, and mutation in a diploid strain of S. cerevisiae. We demonstrate that inducing doses of UV irradiation followed by a 6 h period of incubation render the cells resistant to challenge doses of UV irradiation. The effects of inducing and challenge doses of UV irradiation upon interchromosomal gene conversion and mutation are strictly additive. Using the yeast URA3 gene cloned in non-replicating single- and double-stranded plasmid vectors that integrate into chromosomal genes upon transformation, we show that UV irradiation of haploid yeast cells and homologous plasmid DNA sequences each stimulate homologous recombination approximately two-fold, and that these effects are additive. Non-specific DNA damage has little effect on the stimulation of, homologous recombination, as shown by studies in which UV-irradiated heterologous DNA was included in transformation/recombination experiments. We further demonstrate that the effect of competing single- and double-stranded heterologous DNA sequences differs in UV-irradiated and unirradiated cells, suggesting an induction of recombinational machinery in UV-irradiated S. cerevisiae cells.
Similar content being viewed by others
References
Cox BS, Game JC (1974) Repair systems in Saccharomyces. Mutat Res 20:257–264
Cox MM, Lehman JR (1981) RecA protein of Escherichia coli promotes branch migration, a kinetically distinct phase of DNA strand exchange. Proc Natl Acad Sci USA 78:3433–3437
Elledge SJ, Davis RW (1987) Identification of the gene encoding the small subunit of ribonucleotide reductase from Saccharomyces cerevisiae: DNA damage-inducible gene required for mitotic viability. Mol Cell Biol 7:2783–2893
Fabre F, Roman H (1977) Genetic evidence for inducibility of recombination competence in yeast. Proc Natl Acad Sci USA 74:1667–1671
Falco SC, Rose M, Botstein D (1983) Homologous recombination between episomal plasmids and chromosomes in yeast. Genetics 105:843–856
Friedberg EC (1988) Deoxyribonucleic acid repair in the yeast Saccharomyces cerevisiae. Microbiol Rev 52:70–102
Hinnen A, Hicks JB, Fink GR (1978) Transformation of yeast. Proc Natl Acad Sci USA 75:1929–1933
Holliday R (1971) Biochemical measure of the time and frequency of radiation-induced allelic recombination in Ustilago maydis. Nature New Biol 232:233–236
Holliday R (1975) Further evidence for an inducible recombination repair system in Ustilago maydis. Mutat Res 29:149–153
Jinks-Robertson S, Petes TD (1985) High-frequency meiotic gene conversion between repeated genes on non-homologous chromosomes in yeast. Proc. Natl Acad Sci USA 82:3350–3354
Kunz BA, Haynes RH (1981) Phenomenology and genetic control of mitotic recombination in yeast. Annu Rev Genet 15:57–89
Lee, MG, Yarranton GT (1982) Inducible DNA repair in Ustilago maydis. Mol Gen Genet 185:245–250
Little JW, Mount DW (1982) The SOS regulatory system of Escherichia coli. Cell 29:11–22
Little JW, Edmiston SH, Parcelli LZ, Mount DW (1980) Cleavage of the E. coli lexA protein by the RecA protease. Proc Natl Acad Sci USA 77:3225–3229
Maga JA, McEntee K (1985) Response of S. cerevisiae to N-methyl-N′-nitro-N-nitrosoguanidine: mutagenesis, survival and DDR gene expression. Mol Gen Genet 200:313–320
Maloney DH, Fogel S (1980) Mitotic recombination in yeast: isolation and characterization of mutants with enhanced spontaneous mitotic gene conversion rates. Genetics 94:825–839
McClanahan T, McEntee K (1984) Specific, transcripts are elevated in Saccharomyces cerevisiae in response to DNA damage. Mol Cell Biol 4:2356–2363
Messing J (1983) New M13 vectors for cloning. Methods Enzymol 101:20–79
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
Prakash L (1981) Characterization of postreplication repair in Saccharomyces cerevisiae and effects of rad16, rad18, rev3 and rad52 mutations. Mol Gen Genet 184:471–478
Resnick MA (1976) The repair of double-strand breaks in DNA: A model involving recombination. J Theor Biol 59:97–106
Resnick MA, Martin P (1976) The repair of double-strand breaks in the nuclear DNA of Saccharomyces cerevisiae and its genetic control. Mol Gen Genet 143:119–129
Resnick MA, Stasiewicz S, Game JC (1983) Meiotic DNA metabolism in wild-type and excision-deficient yeast following UV exposure. Genetics 104:583–601
Robinson GW, Nicolet CM, Kalaninov D, Friedberg EC (1986) A yeast excision-repair gene is inducible by DNA damaging agents. Proc Natl Acad Sci USA 83:1842–1846
Roman H, Jacob F (1958) A comparison of spontaneous and ultraviolet-induced allelic recombination with reference to the recombination of outside markers. Cold Spring Harbor Symp Quant Biol 23:155–160
Rose M, Grisafi P, Botstein D (1984) Structure and function of the yeast URA3 gene: expression in Escherichia coli. Gene 29:113–124
Rose M, Winston F (1984) Identification of a Ty insertion within the coding sequence of the S. cerevisiae URA3 gene. Mol Gen Genet 193:557–560
Ruby SW, Szostak JW (1985) Specific Saccharomyces genes are expressed in response to DNA damaging agents. Mol Cell Biol 5:75–84
Salles B, Paoletti C (1983) Control of UV induction of RecA protein. Proc Natl Acad Sci USA 80:65–69
Sherman F, Fink GR, Hicks JB (1986) Methods in yeast genetics. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York
Simon JR, Moore PD (1987) Homologous recombination between single-stranded DNA and chromosomal genes in Saccharomyces cerevisiae. Mol Cell Biol, 7:2329–2334
Unrau P, Wheatcroft R, Cox BS, Olive T (1973) The formation of pyrimidine dimers in the DNA of fungi and bacteria. Biochim Biophys Acta 312:626–632
White CI, Sedgwick SG (1985) The use of plasmid DNA to probe DNA repair functions in the yeast Saccharomyces cerevisiae. Mol Gen Genet 201:99–106
Wilkie D, Lewis D (1963) The effect of ultraviolet light on recombination in yeast. Genetics 48:1701–1716
Zimmermann FK, Kern R, Rasenberger H (1975) A yeast strain for simultaneous detection of induced mitotic crossing over, mitotic gene conversion, and reverse mutation. Mutat Res 28:381–388
Author information
Authors and Affiliations
Additional information
Communicated by G.R. Smith
Rights and permissions
About this article
Cite this article
Simon, J.R., Moore, P.D. Induction of homologous recombination in Saccharomyces cerevisiae . Mol Gen Genet 214, 37–41 (1988). https://doi.org/10.1007/BF00340176
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00340176