Probing the chromosome 9p21 region susceptible to DNA double-strand breaks in human cells in vivo by restriction enzyme transfer

Sato, Masanori; Sasaki, Hiroki; Kazui, Teruhisa; Yokota, Jun

doi:10.1038/sj.onc.1208750

Cited by 4 publications

(2 citation statements)

References 41 publications

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“…The difference in the kinetics of Ser186 phosphorylation upon UV and IR might be due to difference in the nature of IRand UV-induced DNA damage. IR causes single-strand breaks (SSBs), and if the breaks are localized closely to each other in opposite strands, DSBs also occur (Sato et al 2005). The Mre11, NBS1, and Rad50 are critical for the processing of DNA ends damaged by IR and for the formation of RPA-coated ssDNA (Jazayeri et al 2005;Zhong et al 2005;Myers and Cortez 2006).…”

Section: Atr/atm-mediated Phosphorylation Of Cep164mentioning

confidence: 99%

Cep164 is a mediator protein required for the maintenance of genomic stability through modulation of MDC1, RPA, and CHK1

Sivasubramaniam¹,

Sun²,

Pan³

et al. 2008

Genes Dev.

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The activation of the ataxia telangiectasia mutated (ATM) and ATM/Rad3-related (ATR) kinases triggers a diverse cellular response including the initiation of DNA damage-induced cell cycle checkpoints. Mediator of DNA Damage Checkpoint protein, MDC1, and H2AX are chromatin remodeling factors required for the recruitment of DNA repair proteins to the DNA damage sites. We identified a novel mediator protein, Cep164 (KIAA1052), that interacts with both ATR and ATM. Cep164 is phosphorylated upon replication stress, ultraviolet radiation (UV), and ionizing radiation (IR). Ser186 of Cep164 is phosphorylated by ATR/ATM in vitro and in vivo. The phosphorylation of Ser186 is not affected by RPA knockdown but is severely hampered by MDC1 knockdown. siRNA-mediated silencing of Cep164 significantly reduces DNA damage-induced phosphorylation of RPA, H2AX, MDC1, CHK2, and CHK1, but not NBS1. Analyses of Cep164 knockdown cells demonstrate a critical role of Cep164 in G2/M checkpoint and nuclear divisions. These findings reveal that Cep164 is a key player in the DNA damage-activated signaling cascade.[Keywords: DNA damage signal pathways; ATR; CEP164; RPA; MDC1; CHK1; H2AX] Supplemental material is available at http://www.genesdev.org. The genome is subjected to continuous damage and repair. Free radicals generated during cellular metabolism, DNA replication errors, and exogenous carcinogens can all lead to DNA damage, which triggers multiple signal transduction pathways to slow down cell cycle progression, allowing for the repair of the damaged DNA. It has been proposed that these pathways sense DNA damage, activate cell cycle checkpoints, and recruit repair proteins to the damaged DNA Stucki and Jackson 2006). ATM (ataxia telangiectasia mutated) and ATR (ATM-and Rad3-related gene) are two evolutionarily conserved phosphatidylinositol kinase-related proteins that play critical roles in checkpoint activation (Shiloh 2003). Mutations in ATM lead to AT (ataxia telangiectasia) disorder (Savitsky et al. 1995), which is characterized by neuronal degeneration and cancer predisposition, while reduced ATR expression leads to Seckel syndrome (O'Driscoll et al. 2003), which is characterized by retarded development. Knockout of ATM and ATR in mice, respectively, demonstrates that ATR, but not ATM, is essential for embryogenesis and cell survival (Barlow et al. 1996;Brown and Baltimore 2000;Cortez et al. 2001).ATM is activated by DNA double-strand break (DSB)-causing agents including ionizing radiation (IR), while ATR is activated by ultraviolet radiation (UV) as well as replication block (Abraham 2001). Interestingly, IR also activates ATR in an ATM-and Mre11-NBS1-Rad50 (MRN)-dependent manner (Jazayeri et al. 2005;Zhong et al. 2005;Myers and Cortez 2006). ATM and ATR share many substrates; i.e., Ser15 of p53 (Siliciano et al. 1997), and Ser1423 and Ser1524 of BRCA1 (Fabbro et al. 2004). In addition, phosphorylation of Ser139 of H2AX by ATM/ATR is well documented (Burma et al. 2001). Mediator of DNA damage checkpoint protein 1, MDC1, is a c...

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Section: Atr/atm-mediated Phosphorylation Of Cep164mentioning

confidence: 99%

Cep164 is a mediator protein required for the maintenance of genomic stability through modulation of MDC1, RPA, and CHK1

Sivasubramaniam¹,

Sun²,

Pan³

et al. 2008

Genes Dev.

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“…Sato et al (2005) reported a target region including CDKN2A and CDKN2B promoters in the HeLa cell line. The cloning of deletion breakpoints in bladder and renal cancer cell lines revealed that deletions were located in, or close to, a LINE-1 retrotransposon clusters (Florl and Schulz, 2003).…”

Section: Discussionmentioning

confidence: 98%

Molecular characterization of 9p21 deletions shows a minimal common deleted region removing CDKN2A exon 1 and CDKN2B exon 2 in diffuse large b‐cell lymphomas

Guney

Bertrand

Jardin

et al. 2011

Genes Chromosomes & Cancer

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Diffuse large B cell lymphoma represent the most frequent subtype of non Hodgkin's lymphoma, accounting for 30-40% of cases. Several studies have shown that CDKN2A and CDKN2B deletions are frequent in these lymphomas. These genes encode the P14ARF, CDKN2A, and CDKN2B proteins which play a key role in the control of the G1/S transition of the cell cycle. Using array CGH and a quantitative multiplex PCR method, we have previously identified such deletions in 36% of cases at diagnosis. Using a walking strategy to approach the breakpoints of these deletions we could identify the breakpoints junction of thirteen deletions in 11 patients and of two unbalanced translocation leading to a loss of these genes. A minimal common deleted region of 22.4 kb containing exon 1β of CDKN2A encoding P14ARF and exon 2 of CDKN2B encoding CDKN2B was present in all cases but one. Analysis by quantitative RT PCR showed that the expression level of these genes was decreased in these patients as compared with non deleted cases and that this level was correlated with the deletion status of P14ARF, CDKN2A, and CDKN2B. Analysis of the breakpoint sequences showed that some of them were clustered within a few hundred base-pairs suggesting, even if we failed to identify any clear recombination prone sequences, that in these deletions the rearrangement results from non-random mechanisms.

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Chromosomal instability in bladder cancer

Florl

Schulz

2008

Arch Toxicol

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Chromosomal instability (CIN) distinguishes invasive urothelial carcinomas from less malignant papillary subtypes. Recent results implicate checkpoint dysfunction as a crucial factor underlying the emergence of aneuploidy in urothelial carcinogenesis. It may moreover contribute to DNA repair defects. Therefore, defects in cell cycle regulation, p53 function, and checkpoint signaling initially caused by carcinogens in the urothelium could ultimately elicit CIN. Among several mechanisms contributing to aneuploidy, breakage-fusion-bridge (BFB) cycles initiated by defective telomeres may be particularly relevant. The mechanism generating large interstitial deletions, prominently at 9p21, appears to be distinct. New experimental approaches are required to address important unresolved questions such as the precise relationship between telomere erosion and telomerase activation, the influence of checkpoint defects on DNA double-strand repair by non-homologous and homomologous recombination repair systems, and the mechanism responsible for megabase-sized interstitial deletions.

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Probing the chromosome 9p21 region susceptible to DNA double-strand breaks in human cells in vivo by restriction enzyme transfer

Cited by 4 publications

References 41 publications

Cep164 is a mediator protein required for the maintenance of genomic stability through modulation of MDC1, RPA, and CHK1

Cep164 is a mediator protein required for the maintenance of genomic stability through modulation of MDC1, RPA, and CHK1

Molecular characterization of 9p21 deletions shows a minimal common deleted region removing CDKN2A exon 1 and CDKN2B exon 2 in diffuse large b‐cell lymphomas

Chromosomal instability in bladder cancer

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