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Uncapped telomeres are recognized by the DSB repair machinery in senescing human cells, resulting in the formation of γ-H2AX, 53BP1, MDC1 and NBS1 foci. By chromatin immunoprecipitation, these foci form at the telomeres, suggesting that uncapped telomeres are recognized as DSBs. Inactivation of the DNA damage checkpoint allows senescent cells to resume cell cycle progression. Senescing is accompanied by phosphorylation of H2AX, SMC1, RAD17, CHK1 and CHK2, which are also phosphorylated in response to ionizing radiation.
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d'Adda di Fagagna F., Reaper P.M., Clay-Farrace L., Fiegler H., Carr P., Von Zglinicki T., Saretzki G., Carter N.P., Jackson S.P. A DNA damage checkpoint response in telomere-initiated senescence. Nature. 426:2003;194-198 Uncapped telomeres are recognized by the DSB repair machinery in senescing human cells, resulting in the formation of γ-H2AX, 53BP1, MDC1 and NBS1 foci. By chromatin immunoprecipitation, these foci form at the telomeres, suggesting that uncapped telomeres are recognized as DSBs. Inactivation of the DNA damage checkpoint allows senescent cells to resume cell cycle progression. Senescing is accompanied by phosphorylation of H2AX, SMC1, RAD17, CHK1 and CHK2, which are also phosphorylated in response to ionizing radiation.
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1 phase. Furthermore, the clustering is impaired in primary fibroblasts from ataxia telangiectasia-like disorder (ATLD) patients with reduced levels of functional Mre11, suggesting that the ability of the Mre11 complex to tether DNA ends is required for the reorganization of break sites.
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1 phase. Furthermore, the clustering is impaired in primary fibroblasts from ataxia telangiectasia-like disorder (ATLD) patients with reduced levels of functional Mre11, suggesting that the ability of the Mre11 complex to tether DNA ends is required for the reorganization of break sites.
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1 cells. We also demonstrate that the Rad52 recombination protein is modified in a cell-cycle-dependent manner with a higher-mobility form appearing during S phase.
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The authors compare the dynamics of Chk2 and Nbs1 recruitment to laser-induced DSBs. Nbs1 becomes more tightly associated with the DSB sites than Chk2, although both proteins in foci are continuously exchanged with unbound protein. Chk2 becomes phosphorylated by ATM at the DSB sites and the subsequent redistribution of phosphorylated Chk2 throughout the nucleus is required for a proper transmission of the checkpoint response.
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Lukas C., Falck J., Bartkova J., Bartek J., Lukas J. Distinct spatiotemporal dynamics of mammalian checkpoint regulators induced by DNA damage. Nat Cell Biol. 5:2003;255-260 The authors compare the dynamics of Chk2 and Nbs1 recruitment to laser-induced DSBs. Nbs1 becomes more tightly associated with the DSB sites than Chk2, although both proteins in foci are continuously exchanged with unbound protein. Chk2 becomes phosphorylated by ATM at the DSB sites and the subsequent redistribution of phosphorylated Chk2 throughout the nucleus is required for a proper transmission of the checkpoint response.
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Lukas, C.1
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Nuclear dynamics of RAD52 group homologous recombination proteins in response to DNA damage
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The authors use fluorescence loss in photobleaching (FLIP) and fluorescence redistribution after photobleaching (FRAP) to study the dynamics of Rad51, Rad52 and Rad54 in Chinese hamster ovary cells. In the absence of DNA damage, the three proteins exhibit different diffusion rates, demonstrating that they are not part of the same multiprotein complex. In response to ionizing radiation, the three proteins form nuclear foci. The Rad54 and Rad52 proteins at foci are freely exchangeable with unbound proteins and their residence times in foci are 0.5s and 26s, respectively. In contrast, Rad51 associated with foci is not exchanged with unbound protein, indicating that it is found at the core of the DNA-damage-induced foci, probably as a filament bound to single-stranded DNA.
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Essers J., Houtsmuller A.B., van Veelen L., Paulusma C., Nigg A.L., Pastink A., Vermeulen W., Hoeijmakers J.H., Kanaar R. Nuclear dynamics of RAD52 group homologous recombination proteins in response to DNA damage. EMBO J. 21:2002;2030-2037 The authors use fluorescence loss in photobleaching (FLIP) and fluorescence redistribution after photobleaching (FRAP) to study the dynamics of Rad51, Rad52 and Rad54 in Chinese hamster ovary cells. In the absence of DNA damage, the three proteins exhibit different diffusion rates, demonstrating that they are not part of the same multiprotein complex. In response to ionizing radiation, the three proteins form nuclear foci. The Rad54 and Rad52 proteins at foci are freely exchangeable with unbound proteins and their residence times in foci are 0.5s and 26s, respectively. In contrast, Rad51 associated with foci is not exchanged with unbound protein, indicating that it is found at the core of the DNA-damage-induced foci, probably as a filament bound to single-stranded DNA.
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-/- and H2AX(S136A/S139A) mouse cells. Interestingly, H2AX phosphorylation is required for the persistence of Nbs1, 53BP1 and BRCA1 foci at later time-points (four hours after damage induction). This observation indicates that γ-H2AX plays an important role in the later steps of DSB repair.
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Celeste, A.1
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This paper shows that adenovirus evades the host DNA damage response by degrading Mre11. The data demonstrate that Mre11 is a key early component of the DNA checkpoint response required for activation of the ATR and ATM checkpoint kinases.
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This work analyzes the activation of ATM as measured by its autophosphorylation after DNA damage in human wild type, NBS, A-TLD(M) and A-TLD(S) cell lines. Cell lines lacking any of the components of the MRN complex are deficient in ATM activation and phosphorylation of downstream ATM targets such as p53, Chk2 and Hdm2. Furthermore, ectopic expression of wild type Mre11, but not of a nuclease-defective mre11(H129L/D130V) mutant in A-TLD(S) cells, fully reconstitutes ATM activation.
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Uziel T., Lerenthal Y., Moyal L., Andegeko Y., Mittelman L., Shiloh Y. Requirement of the MRN complex for ATM activation by DNA damage. EMBO J. 22:2003;5612-5621 This work analyzes the activation of ATM as measured by its autophosphorylation after DNA damage in human wild type, NBS, A-TLD(M) and A-TLD(S) cell lines. Cell lines lacking any of the components of the MRN complex are deficient in ATM activation and phosphorylation of downstream ATM targets such as p53, Chk2 and Hdm2. Furthermore, ectopic expression of wild type Mre11, but not of a nuclease-defective mre11(H129L/D130V) mutant in A-TLD(S) cells, fully reconstitutes ATM activation.
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2+ ion. Mutation of either cysteine confers sensitivity to ionizing radiation. This work provides a structural explanation for the ability of Rad50 to tether DNA molecules, as demonstrated by electron microscopy.
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2+ ion. Mutation of either cysteine confers sensitivity to ionizing radiation. This work provides a structural explanation for the ability of Rad50 to tether DNA molecules, as demonstrated by electron microscopy.
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