Nucleotide Excision Repair- and Polymerase η-Mediated Error-Prone Removal of Mitomycin C Interstrand Cross-Links

Zheng, Huyong; Wang, Xin; Warren, Amy J.; Legerski, Randy J.; Nairn, Rodney S.; Hamilton, Joshua W.; Li, Lei

doi:10.1128/mcb.23.2.754-761.2003

Cited by 135 publications

(199 citation statements)

References 48 publications

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“…The mutagenic repair of triple helix-directed psoralen cross-links in yeast requires PRR and shows a partial dependence on NER, in agreement with the current study (53,54). A pathway for the error-prone repair of cross-links that depends on transcription-coupled NER and the trans-lesion bypass polymerase has been described in mammalian cells (57,58).…”

Section: Discussionsupporting

confidence: 76%

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: Discussionsupporting

confidence: 76%

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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“…However, most prior studies of pol η and ICLs have been performed utilizing plasmid substrates, rather than analyzing genomic DNA [18][19][20]. We confirmed that loss of pol η renders cells moderately hypersensitive to genomic photoadducts of the psoralen derivative, HMT, under split-dose UVA conditions designed to generate significant quantities of ICL.…”

Section: Discussionsupporting

confidence: 61%

“…While both DSB and ssDNA have been identified as potential intermediate products of replication arrested at a DNA lesion and likely account for a γ-H2AX signal during S-phase, ssDNA associated with incomplete NER may account for γ-H2AX generated during other phases of the cell cycle. NER may operate to uncouple one strand of the ICL, followed by pol η-mediated TLS using the remaining intact strand as a template, as has been generally proposed for other polymerases previously [15,16,20]. While a role for gap-filling during NER by pol η at lesions outside of a replication fork has not been specifically described, polymerase κ, another bypass polymerase in the Y-family to which pol η also belongs, has been proposed to function in this role in NER [44].…”

Section: Discussionmentioning

confidence: 99%

“…Although results from yeast suggest that pol η does not confer resistance to ICL by cisplatin [5], XP-V cells have been reported to be hypersensitive to and impaired in the repair of a number of DNA damaging agents, including ICL-forming agents, suggesting that TLS by pol η is important in lesion tolerance and survival following ICL in humans [17,20,32,39]. However, most prior studies of pol η and ICLs have been performed utilizing plasmid substrates, rather than analyzing genomic DNA [18][19][20].…”

Section: Discussionmentioning

confidence: 99%

“…While rad30-deficient yeast lacking pol η are not dramatically more sensitive to some ICL agents [5], studies in XP-V cells have suggested that pol η is important for ICL repair in humans. XP-V cells are hypersensitive to psoralen photoadducts, including ICL [17], exhibit increased mutagenesis in response to psoralen ICL associated with triplex-forming oligonucleotides [18], are moderately impaired in reactivation of psoralen crosslinked reporter plasmids [19], and are defective in recombination-independent ICL repair of mitomycin C-induced ICL in a plasmid reporter reactivation assay [20]. However, almost all of these studies have been limited to studying ICL in non-replicating plasmids rather than in native genomic DNA [18][19][20].…”

Section: Introductionmentioning

confidence: 99%

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DNA polymerase η reduces the γ-H2AX response to psoralen interstrand crosslinks in human cells

Mogi

Butcher

2008

Experimental Cell Research

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DNA interstrand crosslinks are processed by multiple mechanisms whose relationships to each other are unclear. Xeroderma pigmentosum-variant (XP-V) cells lacking DNA polymerase η are sensitive to psoralen photoadducts created under conditions favoring crosslink formation, suggesting a role for translesion synthesis in crosslink repair. Because crosslinks can lead to double strand breaks, we monitored phosphorylated H2AX (γ-H2AX), which is typically generated near double-strand breaks but also in response to single-stranded DNA, following psoralen photoadduct formation in XP-V fibroblasts to assess whether polymerase η is involved in processing crosslinks. In contrast to conditions favoring monoadducts, conditions favoring psoralen crosslinks induced γ-H2AX levels in both XP-V and nucleotide excision repair-deficient XP-A cells relative to control repair-proficient cells; ectopic expression of polymerase η in XP-V cells normalized the γ-H2AX response. In response to psoralen crosslinking, γ-H2AX as well as 53BP1 formed co-incident foci that were more numerous and intense in XP-V and XP-A cells than in controls. Psoralen photoadducts induced γ-H2AX throughout the cell cycle in XP-V cells. These results indicate that polymerase η is important in responding to psoralen crosslinks, and are consistent with a model in which nucleotide excision repair and polymerase η are involved in processing crosslinks and avoiding γ-H2AX associated with double strand breaks and single-stranded DNA in human cells.

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DNAInterstrand Crosslink Repair

Siede¹

2009

Encyclopedia of Life Sciences

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Crosslinking agents such as psoralens, nitrogen mustards or cisplatin are bifunctionally acting chemicals that generate a fraction of their adducts as covalent linkages between complementary deoxyribonucleic acid strands. Since many of these agents are of importance in genetic toxicology and cancer therapy, repair of interstrand crosslinks (ICL) has been studied extensively in prokaryotes and in lower and higher eukaryotes. The main repair pathway in E. coli involves the sequential action of nucleotide excision repair and recombinational repair. In eukaryotes, several repair pathways play important roles not only in repair including nucleotide excision repair, translesion synthesis, recombination, but also mismatch repair. Relative contributions of the various pathways depend on cell cycle position and agent used. Eukaryotic proteins that specifically enhance resistance to crosslinking agents have been identified ( SNM1 , FANC family of proteins). Key concepts: Chemicals with two or more correctly spaced reactive groups can covalently link opposing DNA strands. Several repair or tolerance pathways such as nucleotide excision repair, recombination and translesion synthesis can work together to overcome such complex damage. Cell cycle stage may determine the choice of repair pathway combinations. A seemingly deleterious consequence of complex DNA damage (replication fork collapse at a crosslinked site) can be an important integrated part of damage tolerance and repair. A heritable human syndrome with multiple diverse phenotypes (Fanconi anaemia) can be associated with a specific DNA repair deficiency (crosslink repair).

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Nucleotide Excision Repair- and Polymerase η-Mediated Error-Prone Removal of Mitomycin C Interstrand Cross-Links

Cited by 135 publications

References 48 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

DNA polymerase η reduces the γ-H2AX response to psoralen interstrand crosslinks in human cells

DNAInterstrand Crosslink Repair

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