Summary
Psoralen photoreaction with DNA produces interstrand crosslinks, which require the activity of excision and recombinational pathways for repair. Yeast replicating plasmids, carrying the HIS3, TRP1, and URA3 genes, were photoreacted with psoralen in vitro and transfected into Saccharomyces cerevisiae cells. Repair was assayed as the relative transformation efficiency. A recombination-deficient rad52 strain was the least efficient in the repair of psoralen-damaged plasmids; excision repair-deficient rad1 and rad3 strains had repair efficiencies intermediate between those of rad52 and RAD cells. The level of repair also depended on the conditions of transformant selection; repair was more efficient in medium lacking tryptophan than in medium from which either histidine or uracil was omitted. The plasmid repair differential between these selective media was greatest in rad1 cells, and depended on RAD52. Plasmid-chromosome recombination was stimulated by psoralen damage, and required RAD52 function. Chromosome to plasmid gene conversion was seen most frequently at the HIS3 locus. In RAD and rad3 cells, the majority of the conversions were associated with plasmid integration, while in rad1 cells most were non-crossover events. Plasmid to chromosome gene conversion was observed most frequently at the TRP1 locus, and was accompanied by plasmid loss.
Similar content being viewed by others
References
Aguilera A, Klein HL (1989) Yeast intrachromosomal recombination: long gene conversion tracts are preferentially associated with reciprocal exchange and require the RAD1 and RAD3 gene products. Genetics 123:683–694
Averbeck D, Moustacchi E (1975) 8-methoxypsoralen plus 365 nm light. Effects and repair in yeast. Biochim Biophys Acta 395:393–404
Averbeck D, Averbeck S, Cundari E (1987) Mutagenic and recombinogenic action of DNA monoadducts photoinduced by the bifunctional furocoumarin 8-methoxypsoralen in yeast (Saccharomyces cerevisiae). Photochem Photobiol 45:371–379
Beggs JD (1978) Transformation of yeast by a replicating hybrid plasmid. Nature 275:104–108
Bohr VA, Wassermann K (1988) DNA repair at the level of the gene. Trends Biochem Sci 13:429–431
Bohr VA, Smith CA, Okumoto DS, Hanawalt PC (1985) DNA repair in an active gene: removal of pyrimidine dimers from the DHFR gene of CHO cells is much more efficient than in the genome overall. Cell 40:359–369
Cassier C, Chanel R, Moustacchi E (1985) Repair of 8-methoxypsoralen photoinduced crosslinks and monoadducts: role of the different repair pathways in yeast. Photochem Photobiol 41:289–294
Chattoraj KK, Cordes K, Berman ML, Das A (1984) Mutagenesis and mutation transfer induced by ultraviolet light in plasmid-cloned DNA. Gene 27:213–222
Friedberg EC (1988) Deoxyribonucleic acid repair in the yeast Saccharomyces cerevisiae. Microbiol Rev 52:70–102
Game JC, Zamb TJ, Braun RJ, Resnick MA, Roth RM (1980) The role of radiation (rad) genes in meiotic recombination in yeast. Genetics 94:51–68
Glaser VM, Glasunov AV, Terzadze GG, Perera JR, Shestakov SV (1990) Genetic control of plasmid DNA double-strand gap repair in yeast. Curr Genet 18:1–5
Hearst JE, Isaacs ST, Kanne D, Rapoport H, Straub K (1984) The reaction of the psoralens with deoxyribonucleic acid. Quart Rev Biophys 171:1–44
Henriques JAP, Moustacchi E (1980) Sensitivity to photoaddition of mono- and bifunctional furocoumarins of X-ray sensitive mutants of Saccharomyces cerevisiae. Photochem Photobiol 31:557–563
Jachymczyk WJ, von Borstel RC, Mowat MRA, Hastings PJ (1981) Repair of interstrand crosslinks in DNA of Saccharomyces cerevisiae requires two systems for DNA repair: the RAD3 system and the RAD51 system. Mol Gen Genet 182:196–205
Jackson JA, Fink GR (1981) Gene conversion between duplicated genetic elements in yeast. Nature 292:306–311
Keil RL, Roeder GS (1984) Cis-acting recombination-stimulating activity in a fragment of the ribosomal DNA in S. cerevisiae. Cell 39:377–386
Klein HL (1988) Different types of recombination events are controlled by the RAD1 and RAD52 genes of Saccharomyces cerevisiae. Genetics 120:367–377
Kunz BA, Haynes RH (1981) Phenomenology and genetic control of mitotic recombination in yeast. Annu Rev Genet 15:57–89
Kunz BA, Kohalmi L, Kang X, Magnusson KA (1990) Specificity of the mutator effect caused by disruption of the RAD1 excision repair gene of Saccharomyces cerevisiae. J Bacteriol 172:3009–3014
Luisi-DeLuca C, Porter RD, Taylor WD (1984) Stimulation of recombination between homologous sequences on plasmid DNA and chromosomal DNA in Escherichia coli by N-acetoxy-2-acetylaminofluorene. Proc Natl Acad Sci USA 81:2831–2835
Magafña-Schwencke N, Henriques JAP, Chanet R, Moustacchi E (1982) The fate of 8-methoxypsoralen photoinduced crosslinks in nuclear and mitochondrial yeast DNA: comparison of wildtype and repair-deficient strains. Proc Natl Acad Sci USA 79:1722–1726
Magaña-Schwencke N, Averbeck D (1991) Repair of exogenous (plasmid) DNA damaged by photoaddition of 8-methoxypsoralen in the yeast Saccharomyces cerevisiae. Mutat Res 251:123–131
Malone R, Esposito ME (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
Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York
Miller RD, Prakash L, Prakash S (1982) Genetic control of excision of Saccharomyces cerevisiae interstrand DNA crosslinks induced by psoralen plus near UV light. Mol Cell Biol 2:939–948
Miller RD, Prakash S; Prakash L (1984) Different effects of PAD genes of Saccharomyces cerevisiae on incisions of interstrand crosslinks and monoadducts in DNA induced by psoralen plus near UV light treatment. Photochem Photobiol 39:349–352
Mink M, Basak AN, Kuntzel H (1990) Restoration of the yeast LEU2 gene by transcriptionally controlled recombination between tandem repeats. Mol Gen Genet 223:107–113
Miozzari G, Niederberger P, Hutter R (1978) Tryptophan biosynthesis in Saccharomyces cerevisiae: control of the flux through the pathway. J Bacteriol 134:48–59
Moerschell RP, Tsunasawa S, Sherman F (1988) Transformation of yeast with synthetic oligonucleotides. Proc Natl Acad Sci USA 85:524–528
Mudgett JS, Taylor WD (1986) Reciprocal and non-reciprocal homologous recombination between Escherichia coli chromosomal DNA and ultraviolet light-irradiated plasmid DNA. Gene 49:235–244
Mudgett JS, Manzella JM, Taylor WD (1990) Homologous recombination and mutagenesis of γ-irradiated plasmid DNA in Escherichia coli host cells. Rad Res 124:57–61
Nickoloff JA, Singer JD, Hoekstra MF, Heffron F (1989) Double-strand breaks stimulate alternative mechanisms of recombination repair. J Mol Biol 207:527–541
Orr-Weaver TL, Szostak JW (1983) Yeast recombination: the association between double-strand gap repair and crossing-over. Proc Natl Acad Sci USA 80:4417–4421
Orr-Weaver T, Szostak JW, Rothstein RJ (1981) Yeast transformation: a model system for the study of recombination. Proc Natl Acad Sci USA 78:6354–6358
Perera JR, Glasunov AV, Glaser VM, Boreiko AV (1988) Repair of double-strand breaks in plasmid DNA in the yeast Saccharomyces cerevisiae. Mol Gen Genet 213:421–424
Prakash S, Prakash L, Burke W, Montelone BA (1980) Effect of the RAD52 gene on recombination in Saccharomyces cerevisiae. Genetics 94:31–50
Ray A, Siddiqi I, Kolodkin AL, Stahl FW (1988) Intrachromosomal gene conversion induced by a DNA double-strand break in Saccharomyces cerevisiae. J Mol Biol 201:247–260
Ronne H, Rothstein R (1988) Mitotic sectored colonies: evidence of heteroduplex DNA formation during direct repeat recombination. Proc Natl Acad Sci USA 85:2696–2700
Rothstein RJ (1983) One-step gene disruption in yeast. Meth Enzymol 101:202–211
Saffran WA, Smith ED, Chan SK (1991) Induction of multiple plasmid recombination in Saccharomyces cerevisiae by psoralen reaction and double strand breaks. Nucleic Acids Res 19:5681–5687
Saffran WA, Cantor CR, Smith ED, Magdi M (1992) Psoralen damage-induced plasmid recombination in Saccharomyces cerevisiae: dependence on RAD1 and RAD52. Mutat Res 274:1–9
Sage E, Moustacchi E (1987) Sequence context effect on 8-methoxypsoralen photobinding to defined DNA fragments. Biochemistry 26:3307–3314
Schiestl RH, Prakash S (1988) RAD1, an excision repair gene of Saccharomyces cerevisiae, is also involved in recombination. Mol Cell Biol 8:3619–3626
Schild D, Calderon IL, Contopoulou R, Mortimer RK (1983) Cloning of yeast recombination repair genes and evidence that several are nonessential genes. In: Friedberg EC, Bridges BA (eds) Cellular responses to DNA damage. Alan R Liss, New York, pp 417–427
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 (1990) Transformation and recombination in rad mutants of Saccharomyces cerevisiae. Mol Gen Genet 223:241–248
Stinchcomb DT, Thomas M, Kelly J, Selker E, Davis RW (1980) Eukaryotic DNA segments capable of autonomous replication in yeast. Proc Natl Acad Sci USA 77:4559–4563
Struhl K (1985) Nucleotide sequence and transcriptional mapping of the yeast pet56-his3-ded1 gene region. Nucleic Acids Res 13:8587–8601
Terleth C, van Sluis CA, van de Putte P (1989) Differential repair of UV damage in Saccharomyces cerevisiae. Nucleic Acids Res 17:4433–4439
Thomas BJ, Rothstein R (1989) Elevated recombination rates in transcriptionally active DNA. Cell 56:619–630
Vos JMH, Hanawalt PC (1987) Processing of psoralen adducts in an active human gene: repair and replication of DNA containing monoadducts and interstrand cross-links. Cell 50:789–799
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
Author information
Authors and Affiliations
Additional information
Communicated by R. Devoret
Rights and permissions
About this article
Cite this article
Han, EK., Saffran, W.A. Differential repair and recombination of psoralen damaged plasmid DNA in Saccharomyces cerevisiae . Molec. Gen. Genet. 236, 8–16 (1992). https://doi.org/10.1007/BF00279637
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00279637