Differential repair and recombination of psoralen damaged plasmid DNA in Saccharomyces cerevisiae

Han, Eun-Kyoung; Saffran, Wilma A.

doi:10.1007/bf00279637

Cited by 4 publications

(4 citation statements)

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“…In studies where both RAD] and RAD3 function were examined, their properties differed. In both spontaneous recombination of an inverted repeat (13), and psoralen damageinduced recombination of a replicating plasmid (16) radl strains had a lower percentage of crossing over than RAD or rad3 strains. The RADI gene product Radlp, in a complex with RadlOp, has a single strand DNA endonuclease activity (40) which may function in the generation of reciprocal exchange products.…”

Section: Discussionmentioning

confidence: 99%

“…H32, WS41, WS1O1 and WS5 were derived from W303 by disruption of the RADJ, RAD4, RADIO and RAD52 genes, respectively. WS1-4C is MATa rad3-2 trpl-1 ade2-1 ura3-1 his3-11,15 (16). Plasmid pUC18-HIS3 consists of HIS3 inserted into the BamHI site of pUC18; it does not replicate in yeast cells.…”

Section: Methodsmentioning

confidence: 99%

“…Plasmid pUC18-HIS3 consists of HIS3 inserted into the BamHI site of pUC18; it does not replicate in yeast cells. Plasmid YRpHUT carries HIS3, URA3 and TRPl-ARSI cloned into pBR322 (16). Plasmid DNA was prepared form E. coli DH5a cells by alkaline lysis and purified by CsCl gradients (17).…”

Section: Methodsmentioning

confidence: 99%

“…Plasmid DNA was photoreacted at 350 nm with [3H]4'aminomethyl-4,5',8-trimethylpsoralen (AMT) as previously described, to produce a mixture of crosslinks and monoadducts (16). For the production of psoralen monoadducts by photoreaction at 390 nm, 60 tg/ml pUC18-HIS3 DNA and 5 ,uM [3H]AMT (specific activity 300 mCi/mmol) in 10 mM Tris-HCl, 1 mM EDTA, pH 8.0 (TE buffer), in a total volume of 1.0 ml, were incubated in the dark for 30 minutes.…”

Section: Plasmid Dna Reactionsmentioning

confidence: 99%

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Single strand and double strand DNA damage-induced reciprocal recombination in yeast. Dependence on nucleotide excision repair andRAD1recombination

et al. 1994

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Single strand and double strand DNA damage-induced recombination were compared in the yeast Saccharomyces cerevisiae. The non-replicating plasmid pUC18-HIS3 was damaged in vitro and introduced into yeast cells; plasmid-chromosome recombinants were selected as stable His+ transformants. Single strand damage was produced by UV irradiation at 254 nm or by psoralen photoreaction at 390 nm. Double strand damage was produced by psoralen photoreaction at 350 nm or by restriction endonuclease digestion. Recombinants were classified as resulting from gene conversion without crossing over, single plasmid integration, or multiple plasmid integration. Single and double strand DNA damage produced different patterns of recombination. In repair proficient cells double strand damage induced primarily multiple plasmid integrations, while single strand damage induced higher proportions of gene conversions and single integrations. Reciprocal recombination depended on the RAD1 gene, which is involved in both excision repair and recombination; plasmid integration induced by all forms of damage was decreased in a rad1 disruption strain. Mutation of the RAD3 excision repair gene decreased plasmid integration induced by far UV irradiation and psoralen crosslinks, but not by double strand breaks, which are not substrates of nucleotide excision repair. Double strand break-induced plasmid integration was also decreased by disruption of RAD10, which forms a complex with RAD1; disruption of RAD4 had no effect. Thus, while nucleotide excision repair genes are involved in the processing of damaged DNA to generate recombination intermediates, RAD1 and RAD10 are additionally involved in reciprocal exchange.

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Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Plasmid Dna Reactionsmentioning

confidence: 99%

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Single strand and double strand DNA damage-induced reciprocal recombination in yeast. Dependence on nucleotide excision repair andRAD1recombination

et al. 1994

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Role of homologous recombination in DNA interstrand crosslink repair

Hinz

2010

Environ and Mol Mutagen

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Homologous recombination repair (HRR) encompasses mechanisms that employ homologous DNA sequences as templates for repair or tolerance of a wide range of DNA lesions that inhibit DNA replication in S phase. Arguably the most imposing of these DNA lesions is that of the interstrand crosslink (ICL), consisting of a covalently attached chemical bridge between opposing DNA strands. ICL repair requires the coordinated activities of HRR and a number of proteins from other DNA repair and damage response systems, including nucleotide excision repair, base excision repair, mismatch repair, and translesion DNA synthesis (TLS). Interestingly, different organisms favor alternative methods of HRR in the ICL repair process. E. coli perform ICL repair using a homology-driven damage bypass mechanism analogous to daughter strand gap repair. Eukaryotes from yeast to humans initiate ICL repair primarily during DNA replication, relying on HRR activity to restart broken replication forks associated with double-strand break intermediates induced by nucleolytic activities of other excision repair factors. Higher eukaryotes also employ several additional factors, including members of the Fanconi anemia damage-response network, which further promote replication-associated ICL repair through the activation and coordination of various DNA excision repair, TLS, and HRR proteins. This review focuses on the proteins and general mechanisms of HRR associated with ICL repair in different model organisms.

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Recombinational and Mutagenic Repair of Psoralen Interstrand Cross-links in Saccharomyces cerevisiae

Greenberg

Alberti

Hearst

et al. 2001

Journal of Biological Chemistry

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Psoralen photoreacts with DNA to form interstrand cross-links, which can be repaired by both nonmutagenic nucleotide excision repair and recombinational repair pathways and by mutagenic pathways. In the yeast Saccharomyces cerevisiae, psoralen cross-links are processed by nucleotide excision repair to form double-strand breaks (DSBs). In yeast, DSBs are repaired primarily by homologous recombination, predicting that cross-link and DSB repair should induce similar recombination end points. We compared psoralen cross-link, psoralen monoadduct, and DSB repair using plasmid substrates with site-specific lesions and measured the patterns of gene conversion, crossing over, and targeted mutation. Psoralen cross-links induced both recombination and mutations, whereas DSBs induced only recombination, and monoadducts were neither recombinogenic nor mutagenic. Although the cross-link-and DSB-induced patterns of plasmid integration and gene conversion were similar in most respects, they showed opposite asymmetries in their unidirectional conversion tracts: primarily upstream from the damage site for cross-links but downstream for DSBs. Cross-links induced targeted mutations in 5% of the repaired plasmids; all were base substitutions, primarily T 3 C transitions. The major pathway of psoralen cross-link repair in yeast is error-free and involves the formation of DSB intermediates followed by homologous recombination. A fraction of the cross-links enter an error-prone pathway, resulting in mutations at the damage site.DNA interstrand cross-linkers are used widely in cancer chemotherapy because of their high cytotoxicity in replicating cells (1). These lesions are complex, and their repair involves several different DNA repair pathways. As with other forms of chemical damage, excision repair systems incise the damaged DNA strands; however, there is no undamaged strand to act as a template, and full repair requires the participation of additional pathways. Recombinational repair pathways are involved in restoring the intact duplex structure after excision (2-7). Additionally, cross-links efficiently induce mutations, implicating error-prone pathways in their repair (8,9).Psoralens are photoreactive DNA cross-linking agents that react with pyrimidine bases on opposite DNA strands in the presence of near ultraviolet light; 5Ј-TpA-3Ј sequences are preferred cross-linking sites (10, 11). There are two photoreactive positions in the psoralen molecule, the 4Ј,5Ј furan and the 3,4 pyrone double bonds, which can undergo sequential photoreactions to form cross-links. The major products of the first photoreaction step are furan-side monoadducts; these can undergo a second photoreaction at the pyrone side to generate interstrand cross-links. Psoralen plus ultraviolet A (PUVA) therapy is used to treat the skin disorders psoriasis and vitiligo; although effective, this treatment has been found to induce nonmelanoma skin cancers in a dose-dependent manner (12).Both nucleotide excision repair (NER) 1 and recombinational repair pathways partic...

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Differential repair and recombination of psoralen damaged plasmid DNA in Saccharomyces cerevisiae

Cited by 4 publications

References 49 publications

Single strand and double strand DNA damage-induced reciprocal recombination in yeast. Dependence on nucleotide excision repair andRAD1recombination

Single strand and double strand DNA damage-induced reciprocal recombination in yeast. Dependence on nucleotide excision repair andRAD1recombination

Role of homologous recombination in DNA interstrand crosslink repair

Recombinational and Mutagenic Repair of Psoralen Interstrand Cross-links in Saccharomyces cerevisiae

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