Most hydrogen peroxide-induced histone H2AX phosphorylation is mediated by ATR and is not dependent on DNA double-strand breaks

Katsube, Takanori; Mori, Masataka; Tsuji, Hideo; Shiomi, Tadahiro; Wang, Bing; Liu, Qiang; Nenoi, Mitsuru; Onoda, Makoto

doi:10.1093/jb/mvu021

Cited by 64 publications

(59 citation statements)

References 37 publications

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“…37 Mounting evidence has shown that the ATR kinase is activated by not only DNA damages or replication stress but also various cellular stressors including hypoxia, mechanical stress, and oxidative stress, without evidence of DNA breaks. [38][39][40] Taken together, we suggest that ATM and ATR pathways may transduce distinct stress signals and lead to different cellular outcomes in VSMCs. 41 Currently, it is not clear how CX-5461 activates the ATM/ATR pathway.…”

Section: Discussionmentioning

confidence: 73%

Therapeutic Targeting of RNA Polymerase I With the Small-Molecule CX-5461 for Prevention of Arterial Injury–Induced Neointimal Hyperplasia

Pang

Zhang

et al. 2017

ATVB

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Objective-RNA polymerase I (Pol I)-dependent rRNA synthesis is a determinant factor in ribosome biogenesis and thus cell proliferation. The importance of dysregulated Pol I activity in cardiovascular disease, however, has not been recognized. Here, we tested the hypothesis that specific inhibition of Pol I might prevent arterial injury-induced neointimal hyperplasia. Approach and Results-CX-5461 is a novel selective Pol I inhibitor. Using this tool, we demonstrated that local inhibition of Pol I blocked balloon injury-induced neointima formation in rat carotid arteries in vivo. Neointimal development was associated with augmented rDNA transcriptional activity as evidenced by the increased phosphorylation of upstream binding factor-1. The beneficial effect of CX-5461 was mainly mediated by inducing G2/M cell cycle arrest of proliferating smooth muscle cells without obvious apoptosis. CX-5461 did not induce p53 stabilization but increased p53 phosphorylation and acetylation and activated the ataxia telangiectasia mutated/ataxia telangiectasia and Rad3-related (ATR) pathway. Inhibition of ATR, but not of ataxia telangiectasia mutated, abolished the cytostatic effect of CX-5461 and p53 phosphorylation. In addition, inhibition of p53 or knockdown of the p53 target GADD45 mimicked the effect of ATR inhibition. In vivo experiments showed that the levels of phospho-p53 and acetyl-p53, and activity of the ataxia telangiectasia mutated/ATR pathway were all augmented in CX-5461-treated vessels. Conclusions-Pol

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Section: Discussionmentioning

confidence: 73%

Therapeutic Targeting of RNA Polymerase I With the Small-Molecule CX-5461 for Prevention of Arterial Injury–Induced Neointimal Hyperplasia

Pang

Zhang

et al. 2017

ATVB

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“…A plausible explanation for why the ATM/Chk2 axis was unaffected by Rac1 downregulation in HeLa cells compared to MCF7 cells may be their different p53 levels-low in HeLa cells and high in MCF cells [57]. Additionally, the oxidative stress generated by low doses of gamma and UV treatments used on our cells can directly lead to H2AX phosphorylation in an ATR-dependent and ATM-independent manner (i.e., without DSBs), as observed in the colon adenocarcinoma cell line HCT116 (normal p53 status) [58].…”

Section: Discussionmentioning

confidence: 79%

Rac1 GTPase-deficient HeLa cells present reduced DNA repair, proliferation, and survival under UV or gamma irradiation

et al. 2015

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Rac1 GTPase controls essential cellular functions related to the cytoskeleton, such as motility and adhesion. Rac1 is overexpressed in many tumor cells, including breast cancers, where it is also involved in the proliferation and checkpoint control necessary for the cell's recovery after exposure to ionizing radiation. However, its role in DNA damage and repair remains obscure in other tumor cells and under different genotoxic conditions. Here, we compare HeLa cells with mutants exogenously expressing a dominant-negative Rac1 (HeLa-Rac1-N17) by their responses to DNA damage induced by gamma or UV radiation. In HeLa cells, these treatments led to increased levels of active Rac1 (Rac1-GTP) and of stress fibers, with a diminished ability to migrate compared to untreated cells. However, the reduction of Rac1-GTP in Rac1-N17-deficient clones resulted in much higher levels of polymerized stress fibers accompanied by a strong impairment of cell migration, even after both radiation treatments. With regard to proliferation and genomic stability, dominant-negative Rac1 cells were more sensitive to gamma and UV radiation, exhibiting reduced proliferation and survival consistent with increased DNA damage and delayed or reduced DNA repair observed in this Rac1-deficient clone. The DNA damage response, as indicated by pH2AX and pChk1 levels, was increased in HeLa cells but was not effectively triggered in the Rac1-N17 clone after radiation treatment, which is likely the main cause of DNA damage accumulation. These data suggest that Rac1 GTPase plays an important role in signaling and contributes to the sensitivity of cervical cancer cells under UV or gamma radiation treatments.

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“…The levels of 8-oxodeoxyguanosine (8-oxo-dG) was measured as a marker of H 2 O 2 -mediated DNA damage [35]. Cells were treated with H 2 O 2 , fixed, labeled with an anti-8-oxo-dG antibody and observed by flow cytometry.…”

Section: Resultsmentioning

confidence: 99%

“…γ-H2AX acts as a scaffold for many proteins during the DNA damage response (DDR) and it governs the persistence of the DDR complex [34, 35]. ATM and Rad3 related (ATR) phosphorylates γH2AX during S phase in response to SSB induced replication fork stalling [36, 37].…”

Section: Resultsmentioning

confidence: 99%

BRCA1 and BRCA2 protect against oxidative DNA damage converted into double-strand breaks during DNA replication

et al. 2015

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BRCA1 and BRCA2 mutation carriers are predisposed to develop breast and ovarian cancers, but the reasons for this tissue specificity are unknown. Breast epithelial cells are known to contain elevated levels of oxidative DNA damage, triggered by hormonally driven growth and its effect on cell metabolism. BRCA1- or BRCA2-deficient cells were found to be more sensitive to oxidative stress, modeled by treatment with patho-physiologic concentrations of hydrogen peroxide. Hydrogen peroxide exposure leads to oxidative DNA damage induced DNA double strand breaks (DSB) in BRCA-deficient cells causing them to accumulate in S-phase. In addition, after hydrogen peroxide treatment, BRCA deficient cells showed impaired Rad51 foci which are dependent on an intact BRCA1-BRCA2 pathway. These DSB resulted in an increase in chromatid-type aberrations, which are characteristic for BRCA1 and BRCA2-deficient cells. The most common result of oxidative DNA damage induced processing of S-phase DSB is an interstitial chromatid deletion, but insertions and exchanges were also seen in BRCA deficient cells. Thus, BRCA1 and BRCA2 are essential for the repair of oxidative DNA damage repair intermediates that persist into S-phase and produce DSB. The implication is that oxidative stress plays a role in the etiology of hereditary breast cancer.

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Most hydrogen peroxide-induced histone H2AX phosphorylation is mediated by ATR and is not dependent on DNA double-strand breaks

Cited by 64 publications

References 37 publications

Therapeutic Targeting of RNA Polymerase I With the Small-Molecule CX-5461 for Prevention of Arterial Injury–Induced Neointimal Hyperplasia

Therapeutic Targeting of RNA Polymerase I With the Small-Molecule CX-5461 for Prevention of Arterial Injury–Induced Neointimal Hyperplasia

Rac1 GTPase-deficient HeLa cells present reduced DNA repair, proliferation, and survival under UV or gamma irradiation

BRCA1 and BRCA2 protect against oxidative DNA damage converted into double-strand breaks during DNA replication

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