Stochastic and reversible assembly of a multiprotein DNA repair complex ensures accurate target site recognition and efficient repair

Luijsterburg, Martijn S.; Bornstaedt, Gesa von; Gourdin, Audrey M.; Politi, Antonio Z.; Moné, Martijn J.; Warmerdam, Daniël O.; Goedhart, Joachim; Vermeulen, Wim; Driel, Roel van; Höfer, Thomas

doi:10.1083/jcb.200909175

Cited by 115 publications

(144 citation statements)

References 80 publications

(178 reference statements)

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“…The intra-strand crosslinks induced by platinum-based therapeutic agents are mainly processed by nucleotide excision repair (NER), which is carried out by a minimal set of proteins including XPA, XPC-RAD23B, XPG, RPA, ERCC1-XPF, TFIIH, PCNA, DNA polymerase δ or ε, and DNA ligase I, in human cells [9,10] . These proteins are sequentially assembled at the site of DNA lesions in chromatin [11][12][13] , where they carry out the dual incision of the damagecontaining DNA. Approximately 24-32 nucleotides are then removed, and a new fragment is synthesized to restore the DNA structure [14][15][16] .…”

Section: Repair Of Intra-strand Crosslinksmentioning

confidence: 99%

DNA damage responses in cancer stem cells: Implications for cancer therapeutic strategies

Wang

2015

WJBC

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The identification of cancer stem cells (CSCs) that are responsible for tumor initiation, growth, metastasis, and therapeutic resistance might lead to a new thinking on cancer treatments. Similar to stem cells, CSCs also display high resistance to radiotherapy and chemotherapy with genotoxic agents. Thus, conventional therapy may shrink the tumor volume but cannot eliminate cancer. Eradiation of CSCs represents a novel therapeutic strategy. CSCs possess a highly efficient DNA damage response (DDR) system, which is considered as a contributor to the resistance of these cells from exposures to DNA damaging agents. Targeting of enhanced DDR in CSCs is thus proposed to facilitate the eradication of CSCs by conventional therapeutics. To achieve this aim, a better understanding of the cellular responses to DNA damage in CSCs is needed. In addition to the protein kinases and enzymes that are involved in DDR, other processes that affect the DDR including chromatin remodeling should also be explored.

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Section: Repair Of Intra-strand Crosslinksmentioning

confidence: 99%

DNA damage responses in cancer stem cells: Implications for cancer therapeutic strategies

Wang

2015

WJBC

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“…Because NER can produce ssDNA gaps of ∼30 nt for a variety of bulky lesions, there is a greater likelihood of secondary generation of DSBs than with base-excision repair, which generates short resection regions. However, gap formation and subsequent refilling during NER are tightly coordinated, with repair synthesis starting after incision on the 5′ side of the lesion (which precedes the 3′ incision), resulting in rapid completion of repair (11,12). This coordination might prevent the conversion of ssDNA gaps into more deleterious DSBs, thereby preserving genome stability.…”

mentioning

confidence: 99%

Homologous recombination rescues ssDNA gaps generated by nucleotide excision repair and reduced translesion DNA synthesis in yeast G2 cells

Westmoreland

Resnick

2013

Proc. Natl. Acad. Sci. U.S.A.

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Repair of DNA bulky lesions often involves multiple repair pathways such as nucleotide-excision repair, translesion DNA synthesis (TLS), and homologous recombination (HR). Although there is considerable information about individual pathways, little is known about the complex interactions or extent to which damage in single strands, such as the damage generated by UV, can result in double-strand breaks (DSBs) and/or generate HR. We investigated the consequences of UV-induced lesions in nonreplicating G2 cells of budding yeast. In contrast to WT cells, there was a dramatic increase in ssDNA gaps for cells deficient in the TLS polymerases η (Rad30) and ζ (Rev3). Surprisingly, repair in TLS-deficient G2 cells required HR repair genes RAD51 and RAD52, directly revealing a redundancy of TLS and HR functions in repair of ssDNAs. Using a physical assay that detects recombination between circular sister chromatids within a few hours after UV, we show an approximate three-fold increase in recombinants in the TLS mutants over that in WT cells. The recombination, which required RAD51 and RAD52, does not appear to be caused by DSBs, because a dose of ionizing radiation producing 20 times more DSBs was much less efficient than UV in producing recombinants. Thus, in addition to revealing TLS and HR functional redundancy, we establish that UV-induced recombination in TLS mutants is not attributable to DSBs. These findings suggest that ssDNA that might originate during the repair of closely opposed lesions or of ssDNA-containing lesions or from uncoupled replication may drive recombination directly in various species, including humans.

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“…Several lines of evidence lead to the currently accepted hypothesis that the lesions are initially recognized by XPC with the help of UV-DDB (UV DNA damage binding protein) for cyclobutane pyrimidine dimer (CPD) lesions (17)(18)(19)(20) followed by damage verification by XPD/TFIIH (21)(22)(23)(24), XPA, and XPG (3) to finally assemble to the preincision complex (3,(25)(26)(27). This step seems to involve recruitment of the XPA protein, which is one of the proteins essential for both GG-NER and TC-NER (28).…”

mentioning

confidence: 99%

Structural insights into the recognition of cisplatin and AAF-dG lesion by Rad14 (XPA)

Koch

Kuper

Gasteiger

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

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Nucleotide excision repair (NER) is responsible for the removal of a large variety of structurally diverse DNA lesions. Mutations of the involved proteins cause the xeroderma pigmentosum (XP) cancer predisposition syndrome. Although the general mechanism of the NER process is well studied, the function of the XPA protein, which is of central importance for successful NER, has remained enigmatic. It is known, that XPA binds kinked DNA structures and that it interacts also with DNA duplexes containing certain lesions, but the mechanism of interactions is unknown. Here we present two crystal structures of the DNA binding domain (DBD) of the yeast XPA homolog Rad14 bound to DNA with either a cisplatin lesion (1,2-GG) or an acetylaminofluorene adduct (AAF-dG). In the structures, we see that two Rad14 molecules bind to the duplex, which induces DNA melting of the duplex remote from the lesion. Each monomer interrogates the duplex with a β-hairpin, which creates a 13mer duplex recognition motif additionally characterized by a sharp 70°DNA kink at the position of the lesion. Although the 1,2-GG lesion stabilizes the kink due to the covalent fixation of the crosslinked dG bases at a 90°angle, the AAF-dG fully intercalates into the duplex to stabilize the kinked structure.

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Stochastic and reversible assembly of a multiprotein DNA repair complex ensures accurate target site recognition and efficient repair

Abstract: Computational modeling and quantitative analysis show that although accumulation of repair complexes can take hours, the individual components rapidly exchange between the nucleoplasm and DNA damage sites.

Cited by 115 publications

References 80 publications

DNA damage responses in cancer stem cells: Implications for cancer therapeutic strategies

DNA damage responses in cancer stem cells: Implications for cancer therapeutic strategies

Homologous recombination rescues ssDNA gaps generated by nucleotide excision repair and reduced translesion DNA synthesis in yeast G2 cells

Structural insights into the recognition of cisplatin and AAF-dG lesion by Rad14 (XPA)

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