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

Greenberg, Ross B.; Alberti, Marie; Hearst, John E.; Chua, Mark A.; Saffran, Wilma A.

doi:10.1074/jbc.m103588200

Cited by 30 publications

(30 citation statements)

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“…Physical analysis confirmed that the difference was due to the direction of gene conversion. In previous studies in repair-proficient cells (30) we observed a bias in the directionality of damage-induced gene conversion within HIS3. DSBs placed in the middle of the HIS3 gene induced more conversion in the 3Ј (downstream) direction than in the 5Ј (upstream) direction.…”

Section: Table V Psoralen Cross-link-induced Mutationsmentioning

confidence: 99%

“…Homologous Recombination-deficient Strains-HR is the major repair pathway for DSB repair in S. cerevisiae (11), and the induction of recombination by interstrand cross-links is thought to be due to DSB-containing intermediates in crosslink repair (1,6,7,30). We examined repair, recombination, and mutation in strains deficient in the HR genes RAD51, RAD52, RAD54, RAD55, and RAD57.…”

Section: Alternate Pathways Of Cross-link Repairmentioning

confidence: 99%

“…Plasmids with psoralen monoadducts or cross-links were prepared as previously described (30). Briefly, a 14-base oligonucleotide primer, 5Ј-CAGGCCGTACGCAG-3Ј, was photoreacted with 4Ј-hydroxymethyl-4,5Ј,8-trimethylpsoralen to produce an adduct at the unique T position.…”

Section: Yeast Strains and Plasmids-yeastmentioning

confidence: 99%

“…The amplification product was also digested with BsiWI to detect mutations at the damage site; phis3-622X yields fragments of 523 and 1052 bp, whereas mutants are refractory to BsiWI digestion. Integrated plasmids were characterized by Southern analysis as previously described (30).…”

Section: Yeast Strains and Plasmids-yeastmentioning

confidence: 99%

“…In a previous study of psoralen photodamage repair in repair-proficient yeast cells (30) we found that interstrand crosslinks placed at a specific site within plasmid molecules induced homologous recombination in the majority of repair events and mutations targeted to the damage site in a smaller fraction of the repaired plasmids. There was no overlap between these endpoints, suggesting that the recombinational and error-prone pathways are independent, alternate mechanisms for interstrand cross-link repair.…”

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confidence: 99%

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DNA Repair Defects Channel Interstrand DNA Cross-links into Alternate Recombinational and Error-prone Repair Pathways

Saffran

Ahmed

Bellevue

et al. 2004

Journal of Biological Chemistry

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The repair of psoralen interstrand cross-links in the yeast Saccharomyces cerevisiae involves the DNA repair groups nucleotide excision repair (NER), homologous recombination (HR), and post-replication repair (PRR). In repair-proficient yeast cells cross-links induce double-strand breaks, in an NER-dependent process; the double-strand breaks are then repaired by HR. An alternate error-prone repair pathway generates mutations at cross-link sites. We have characterized the repair of plasmid molecules carrying a single psoralen cross-link, psoralen monoadduct, or double-strand break in yeast cells with deficiencies in NER, HR, or PRR genes, measuring the repair efficiencies and the levels of gene conversions, crossing over, and mutations. Strains with deficiencies in the NER genes RAD1, RAD3, RAD4, and RAD10 had low levels of cross-link-induced recombination but higher mutation frequencies than repair-proficient cells. Deletion of the HR genes RAD51, RAD52, RAD54, RAD55, and RAD57 also decreased induced recombination and increased mutation frequencies above those of NER-deficient yeast. Strains lacking the PRR genes RAD5, RAD6, and RAD18 did not have any crosslink-induced mutations but showed increased levels of recombination; rad5 and rad6 cells also had altered patterns of cross-link-induced gene conversion in comparison with repair-proficient yeast. Our observations suggest that psoralen cross-links can be repaired by three pathways: an error-free recombinational pathway requiring NER and HR and two PRR-dependent errorprone pathways, one NER-dependent and one NERindependent.DNA interstrand cross-links are complex lesions, highly toxic to cells, and difficult to repair because of the involvement of both strands of the DNA duplex. A single unrepaired interstrand cross-link is sufficient to kill a cell, and multiple DNA repair pathways may be required to complete cross-link removal (1-7).The yeast Saccharomyces cerevisiae has three major DNA repair epistasis groups, all of which are involved in cross-link repair; they are nucleotide excision repair, recombinational repair, and post-replication repair (8). Nucleotide excision repair (NER) 1 is a general system that recognizes bulky and helix-distorting lesions (9, 10). Endonucleases nick the affected DNA strand on both the 5Ј and 3Ј sides of the lesion; after removal of the damaged oligonucleotide, the resulting singlestrand gap is filled in by polymerase activity using the undamaged strand as a template. This mechanism produces error-free repair of single-strand damage. Recombinational repair is the major pathway for repair of double-strand damage, such as double-strand breaks (DSBs) or gaps in yeast (11,12). In homologous recombination (HR) the broken DNA molecule is repaired, and missing genetic information is restored through interactions with intact homologous sequences. This pathway is also non-mutagenic but can result in DNA rearrangements. Post-replication repair (PRR) acts on damage in the context of DNA replication, allowing for bypass of replication fork-...

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Section: Table V Psoralen Cross-link-induced Mutationsmentioning

confidence: 99%

Section: Alternate Pathways Of Cross-link Repairmentioning

confidence: 99%

Section: Yeast Strains and Plasmids-yeastmentioning

confidence: 99%

Section: Yeast Strains and Plasmids-yeastmentioning

confidence: 99%

mentioning

confidence: 99%

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DNA Repair Defects Channel Interstrand DNA Cross-links into Alternate Recombinational and Error-prone Repair Pathways

Saffran

Ahmed

Bellevue

et al. 2004

Journal of Biological Chemistry

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DNADamage: Repair

Heenan

Sancar

2008

Wiley Encyclopedia of Chemical Biology

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Targeting and processing of site‐specific DNA interstrand crosslinks

Vásquez

2010

Environ and Mol Mutagen

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DNA interstrand crosslinks (ICLs) are among the most cytotoxic types of DNA damage, and thus ICL-inducing agents such as cyclophosphamide, melphalan, cisplatin, psoralen and mitomycin C have been used clinically as anti-cancer drugs for decades. ICLs can also be formed endogenously as a consequence of cellular metabolic processes. ICL-inducing agents continue to be among the most effective chemotherapeutic treatments for many cancers; however, treatment with these agents can lead to secondary malignancies, in part due to mutagenic processing of the DNA lesions. The mechanisms of ICL repair have been characterized more thoroughly in bacteria and yeast than in mammalian cells. Thus, a better understanding the molecular mechanisms of ICL processing offers the potential to improve the efficacy of these drugs in cancer therapy. In mammalian cells it is thought that ICLs are repaired by the coordination of proteins from several pathways, including nucleotide excision repair (NER), base excision repair (BER), mismatch repair (MMR), homologous recombination (HR), translesion synthesis (TLS), and proteins involved in Fanconi anemia (FA). In this review, we focus on the potential functions of NER, MMR, and HR proteins in the repair of and response to ICLs in human cells and in mice. We will also discuss a unique approach, using psoralen covalently linked to triplex-forming oligonucleotides to direct ICLs to specific sites in the mammalian genome.

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

Cited by 30 publications

References 57 publications

DNA Repair Defects Channel Interstrand DNA Cross-links into Alternate Recombinational and Error-prone Repair Pathways

DNA Repair Defects Channel Interstrand DNA Cross-links into Alternate Recombinational and Error-prone Repair Pathways

DNADamage: Repair

Targeting and processing of site‐specific DNA interstrand crosslinks

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