Role of homologous recombination in DNA interstrand crosslink repair

Hinz, John M.

doi:10.1002/em.20577

Cited by 59 publications

(50 citation statements)

References 263 publications

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“…[26][27][28] In this study, we analyzed the cell cycle dependence of ACNU, Cell Cycle volume 11 issue 14 junction resolution (XRCC3). 24 The BRCA2, Rad51 and XRCC3 mutants proved to be extremely sensitive to ACNU; toxic effects were already observed with the lowest tested concentrations of 1 and 2.5 μM ACNU (Fig. 4A and B).…”

Section: Experiments With Synchronized Cellsmentioning

confidence: 87%

“…Thus, it has been suggested that once ICL are formed, their repair is complex, utilizing components of nucleotide excision repair, 22 translesion synthesis 23 and DSB repair that operates during the S and G 2 phase of the cell cycle. 24 If CNU-induced lesions, notably O 6 -chloroethylguanine and ICL remain unrepaired, they trigger the cell to activate the apoptotic and necrotic pathway. 25 Previously, it has been shown that methylating agents, such as MNNG and temozolomide, induce cell death triggered by the DNA damage O 6 -methylguanine following the passage through two DNA replication cycles, and that DSB formation and their repair by homologous recombination (HR) and, to a much lesser extent, by nonhomologous end-joining (NHEJ) in the second post-treatment cell cycle is involved.…”

Section: Experiments With Synchronized Cellsmentioning

confidence: 99%

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Chloroethylnitrosourea-induced cell death and genotoxicity: Cell cycle dependence and the role of DNA double-strand breaks, HR and NHEJ

et al. 2012

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C hloroethylnitrosureas (CNUs) are powerful DNA-reactive alkylating agents used in cancer therapy. Here, we analyzed cyto-and genotoxicity of nimustine (ACNU), a representative of CNUs, in synchronized cells and in cells deficient in repair proteins involved in homologous recombination (HR) or nonhomologous end-joining (NHEJ). We show that HR mutants are extremely sensitive to ACNU, as measured by colony formation, induction of apoptosis and chromosomal aberrations. The NHEJ mutants differed in their sensitivity, with Ku80 mutants being moderately sensitive and DNA-PKcs mutated cells being resistant. HR mutated cells displayed a sustained high level of γH2AX foci , which co-stained with Rad51 and 53BP1, indicating DNA double-strand breaks (DSB) to be formed. Using synchronized cells, we analyzed whether DSB formation after ACNU treatment was replication-dependent. We show that γH2AX foci were not induced in G 1 but increased significantly in S phase and remained at a high level in G 2 , where a fraction of cells became arrested and underwent, with a delay of > 12 h, cell death by apoptosis and necrosis. Rad51, ATM, MDC-1 and RPA-2 foci were also formed and shown to co-localize with γH2AX foci induced in S phase, indicating that the DNA damage response was activated. All effects observed were abrogated by MGMT, which repairs O 6 -chloroethylguanine that is converted into DNA cross-links. We deduce that the major genotoxic and killing lesion induced by CNUs are O 6 -chloroethylguanine-triggered

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Section: Experiments With Synchronized Cellsmentioning

confidence: 87%

Section: Experiments With Synchronized Cellsmentioning

confidence: 99%

Chloroethylnitrosourea-induced cell death and genotoxicity: Cell cycle dependence and the role of DNA double-strand breaks, HR and NHEJ

et al. 2012

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“…HDR mutants are characterized by sensitivity to DNA crosslinking agents like mitomycin C (MMC) (27). If primary cells in the animal require HDR, and specifically BRCA1, for repair of cross-links, then BRCA1-deficient mice should be sensitive to these agents.…”

mentioning

confidence: 99%

Double-strand break repair by homologous recombination in primary mouse somatic cells requires BRCA1 but not the ATM kinase

Kass¹,

Helgadottir

Chen

et al. 2013

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

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Homology-directed repair (HDR) is a critical pathway for the repair of DNA double-strand breaks (DSBs) in mammalian cells. Efficient HDR is thought to be crucial for maintenance of genomic integrity during organismal development and tumor suppression. However, most mammalian HDR studies have focused on transformed and immortalized cell lines. We report here the generation of a Direct Repeat (DR)-GFP reporter-based mouse model to study HDR in primary cell types derived from diverse lineages. Embryonic and adult fibroblasts from these mice as well as cells derived from mammary epithelium, ovary, and neonatal brain were observed to undergo HDR at I-SceI endonuclease-induced DSBs at similar frequencies. When the DR-GFP reporter was crossed into mice carrying a hypomorphic mutation in the breast cancer susceptibility gene Brca1, a significant reduction in HDR was detected, showing that BRCA1 is critical for HDR in somatic cell types. Consistent with an HDR defect, Brca1 mutant mice are highly sensitive to the cross-linking agent mitomycin C. By contrast, loss of the DSB signaling ataxia telangiectasia-mutated (ATM) kinase did not significantly alter HDR levels, indicating that ATM is dispensable for HDR. Notably, chemical inhibition of ATM interfered with HDR. The DR-GFP mouse provides a powerful tool for dissecting the genetic requirements of HDR in a diverse array of somatic cell types in a normal, nontransformed cellular milieu.

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“…After filling in the gap, the cross-link remains bound to the second strand undergo excision by NER enzymes. The repair of DSB formed during removal of ICL from the replicating DNA with the participation of FA-proteins, is also provided by the mechanism of HR (Hinz, 2010).…”

Section: Repair Of Interstrand Cross-links Of Dnamentioning

confidence: 99%