Parp-1 protects homologous recombination from interference by Ku and Ligase IV in vertebrate cells

Hochegger, Helfrid; Dejsuphong, Donniphat; Fukushima, Toru; Morrison, Ciarán; Sonoda, Eiichiro; Schreiber, Valérie; Zhao, Guang Yu; Saberi, Alihossein; Masutani, Mitsuko; Adachi, Noritaka; Koyama, Hideki; Murcia, Gilbert de; Takeda, Shunichi

doi:10.1038/sj.emboj.7601015

Cited by 227 publications

(214 citation statements)

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“…PARP-1 has been shown to interact with the NHEJ proteins, Ku antigen and DNA-dependent protein kinase (DNA-PK) with reciprocal modifications and mutual alterations in activity suggested (Ruscetti et al, 1998;Ariumi et al, 1999;Galande and KohwiShigematsu, 1999). Another study indicated that PARP-1 may specifically facilitate HR by decreasing HR-inhibitory effects of Ku antigen (Hochegger et al, 2006). This correlated with a report that the interaction of Ku with PARP-1 modulated the affinity of Ku antigen for DNA ends (Li et al, 2004).…”

Section: Introductionsupporting

confidence: 75%

“…Activation of PARP-1 in response to bleomycin depends on Ku antigen PARP-1 has previously been suggested to have a role in regulating the response of Ku antigen to DSBs PARP-1 activation by bleomycin F Dong et al (Hochegger et al, 2006;Wang et al, 2006). To determine whether Ku might also be important for the activation of PARP-1 subsequent to the DSBs induced by bleomycin, we compared the activity of PARP-1 after treatment in extracts prepared from V79 cells, a hamster embryonic lung fibroblast cell line and V15B cells, a line clonally derived from V79 containing a mutation in the Ku80 gene that leads to the absence of Ku70 and Ku80 (Errami et al, 1996).…”

Section: Resultsmentioning

confidence: 99%

“…Genetic studies have established a role for PARP-1 in HR (Hochegger et al, 2006;Idogawa et al, 2007), whereas biochemical and genetic studies have linked PARP-1 to NHEJ (Schultz et al, 2003;Perrault et al, 2004;Audebert et al, 2006;Wang et al, 2006). PARP-1 has been shown to interact with the NHEJ proteins, Ku antigen and DNA-dependent protein kinase (DNA-PK) with reciprocal modifications and mutual alterations in activity suggested (Ruscetti et al, 1998;Ariumi et al, 1999;Galande and KohwiShigematsu, 1999).…”

Section: Introductionmentioning

confidence: 99%

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Activation of PARP-1 in response to bleomycin depends on the Ku antigen and protein phosphatase 5

Fang¹,

Soubeyrand²,

Haché

2010

Oncogene

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Poly (ADP-ribose) polymerase-1 (PARP-1) has an important role in the cellular response to a broad spectrum of DNA lesions. PARP-1 is strongly activated in response to double-stranded DNA breaks (DSBs), yet its contribution to the DSB response is poorly understood. Here we used bleomycin, a radiomimetic that generates DSBs with high specificity to focus on the response of PARP-1 to DSBs. We report that the induction of PARP-1 activity by bleomycin depends on the Ku antigen, a nonhomologous-DNA-End-Joining factor and protein phosphatase 5 (PP5). PARP-1 activation in response to bleomycin was reduced over 10-fold in Ku-deficient cells, whereas its activation in response to U.V. was unaffected. PARP-1 activation was rescued by reexpression of Ku, but was refractory to manipulation of DNA-dependent protein kinase or ATM. Similarly, PARP-1 activation subsequent to bleomycin was reduced 2-fold on ablation of PP5 and was increased 5-fold when PP5 was overexpressed. PP5 seemed to act directly on PARP-1, as its basal phosphorylation was reduced on overexpression of PP5, and PP5 dephosphorylated PARP-1 in vitro. These results highlight the functional importance of Ku antigen and PP5 for PARP-1 activity subsequent to DSBs.

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Section: Introductionsupporting

confidence: 75%

Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Activation of PARP-1 in response to bleomycin depends on the Ku antigen and protein phosphatase 5

Fang¹,

Soubeyrand²,

Haché

2010

Oncogene

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“…In addition, PARP1 and Ku80 compete for DNA ends in vitro (16). Finally, the genetic ablation of KU70 or LIGIV restores the survival of PARP1-deficient cells exposed to agents inducing DSBs (17,18). These observations raise the question of whether NHEJ is involved in the genomic instability and cytotoxicity observed in HR-deficient cells treated with PARP inhibitors.…”

mentioning

confidence: 72%

Nonhomologous end joining drives poly(ADP-ribose) polymerase (PARP) inhibitor lethality in homologous recombination-deficient cells

Patel

Sarkaria

Kaufmann

2011

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

475

431

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Poly(ADP-ribose) polymerase (PARP) inhibitors are strikingly toxic to cells with defects in homologous recombination (HR). The mechanistic basis for these findings is incompletely understood. Here, we show that PARP inhibitor treatment induces phosphorylation of DNA-dependent protein kinase substrates and stimulates error-prone nonhomologous end joining (NHEJ) selectively in HRdeficient cells. Notably, inhibiting DNA-dependent protein kinase activity reverses the genomic instability previously reported in these cells after PARP inhibition. Moreover, disabling NHEJ by using genetic or pharmacologic approaches rescues the lethality of PARP inhibition or down-regulation in cell lines lacking BRCA2, BRCA1, or ATM. Collectively, our results not only implicate PARP1 catalytic activity in the regulation of NHEJ in HR-deficient cells, but also indicate that deregulated NHEJ plays a major role in generating the genomic instability and cytotoxicity in HR-deficient cells treated with PARP inhibitors.chemotherapy | DNA repair | synthetic lethality | double-strand break repair P oly(ADP-ribose) polymerase 1 (PARP1) is an abundant nuclear enzyme that synthesizes poly(ADP-ribose) polymer when activated by DNA nicks or breaks. Activation of PARP1 has important effects on a variety of cellular processes, including base excision repair (BER), transcription, and cellular bioenergetics (1). The role of PARP1 in the DNA damage response sparked interest in the development of PARP inhibitors as potential chemosensitizers for the treatment of cancer (1, 2). The more recent observation that PARP inhibition is particularly lethal to cells deficient in homologous recombination (HR) proteins (3-8) has generated additional excitement in the cancer chemotherapy community. The current explanation for this hypersensitivity focuses on a mechanism (Fig. 1A) in which loss of PARP1 activity is thought to result in accumulation of DNA single-strand breaks (SSBs), which are subsequently converted to DNA double-strand breaks (DSBs) by the cellular replication and/or transcription machinery. These DSBs, which are repaired by HR in BRCApositive cells, are presumed to accumulate in BRCA1-or BRCA2-deficient cells, leading to subsequent cell death. Heightened sensitivity to PARP inhibition has also been observed in cells with other genetic lesions that affect HR, including phosphatase and tensin homolog (PTEN) deficiency (5), ataxia telangiectasia mutated (ATM) deficiency (7,8), and Aurora A overexpression (6).Although the preceding studies underscore the importance of PARP1 and HR in maintaining genomic stability, they do not address the role of nonhomologous end joining (NHEJ), an alternate DSB repair modality that directly joins broken ends of DNA with little or no regard for sequence homology (9). NHEJ is initiated when free DNA ends are bound by Ku70 and Ku80, which recruit the catalytic subunit of DNA-dependent protein kinase (DNA-PKcs). The resulting complex, known as the DNAdependent protein kinase (DNA-PK) complex, phosphorylates downstream targe...

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“…Moreover, PARP-1 is required for slowing replication fork progression when replication fork collapse as a result of treatment with DSBs inducing agents [23]. The slowing of replication fork progression depends on the recruitment of HR pathway to the damage sites favoured by the ability of PARP-1 to suppress the inhibitory effect of NHEJ on HR [56]. Interestingly, very recent data showed that pharmacological inhibition of PARP activity or silencing of PARP-1 expression down regulates the expression of RAD51 and BRCA1 through the induction of E2F4/p130 transcription factors decreasing the efficacy of the repair mediated by HR [57].…”

Section: Discussionmentioning

confidence: 99%

Common fragile sites in colon cancer cell lines: Role of mismatch repair, RAD51 and poly(ADP-ribose) polymerase-1

Vernole

Muzi

Volpi

et al. 2011

Mutation Research/Fundamental and Molecular Mechanisms of Mutag

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t r a c tCommon fragile sites (CFS) are specific chromosomal areas prone to form gaps and breaks when cells are exposed to stresses that affect DNA synthesis, such as exposure to aphidicolin (APC), an inhibitor of DNA polymerases. The APC-induced DNA damage is repaired primarily by homologous recombination (HR), and RAD51, one of the key players in HR, participates to CFS stability. Since another DNA repair pathway, the mismatch repair (MMR), is known to control HR, we examined the influence of both the MMR and HR DNA repair pathways on the extent of chromosomal damage and distribution of CFS provoked by APC and/or by RAD51 silencing in MMR-deficient and -proficient colon cancer cell lines (i.e., HCT-15 and HCT-15 transfected with hMSH6, or HCT-116 and HCT-116/3+6, in which a part of a chromosome 3 containing the wild-type hMLH1 allele was inserted). Here, we show that MMR-deficient cells are more sensitive to APC-induced chromosomal damage particularly at the CFS as compared to MMR-proficient cells, indicating an involvement of MMR in the control of CFS stability. The most expressed CFS is FRA16D in 16q23, an area containing the tumour suppressor gene WWOX often mutated in colon cancer. We also show that silencing of RAD51 provokes a higher number of breaks in MMR-proficient cells with respect to their MMR-deficient counterparts, likely as a consequence of the combined inhibitory effects of RAD51 silencing on HR and MMR-mediated suppression of HR. The RAD51 silencing causes a broader distribution of breaks at CFS than that observed with APC. Treatment with APC of RAD51-silenced cells further increases DNA breaks in MMR-proficient cells. The RNAi-mediated silencing of PARP-1 does not cause chromosomal breaks or affect the expression/distribution of CFS induced by APC. Our results indicate that MMR modulates colon cancer sensitivity to chromosomal breaks and CFS induced by APC and RAD51 silencing.

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Parp-1 protects homologous recombination from interference by Ku and Ligase IV in vertebrate cells

Cited by 227 publications

References 52 publications

Activation of PARP-1 in response to bleomycin depends on the Ku antigen and protein phosphatase 5

Activation of PARP-1 in response to bleomycin depends on the Ku antigen and protein phosphatase 5

Nonhomologous end joining drives poly(ADP-ribose) polymerase (PARP) inhibitor lethality in homologous recombination-deficient cells

Common fragile sites in colon cancer cell lines: Role of mismatch repair, RAD51 and poly(ADP-ribose) polymerase-1

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