Chromosomal breaks during mitotic catastrophe trigger γH2AX–ATM–p53-mediated apoptosis

Abstract

There was an error published in J. Cell Sci. 124, 2951-2963.In Fig. 4C, the blot representing pATM was inadvertently duplicated as representing G3PDH. The G3PDH western blot has been replaced with the correct image in the figure shown below. There are no changes to the figure legend, which is accurate. This error does not affect the conclusions of the study.The authors apologise to the readers for any confusion that this error might have caused. Fig. 4. Increased γH2AX level during MC leads to cell death. Untreated or doxorubicin (600 nM)-treated 14-3-3σ-knockout HCT116 cells were cultured and harvested at indicated time points and analysed by western blot (50 μg), comet assay and immunostaining using antibodies directed against γH2AX (green). Blots were reprobed for G3PDH to confirm an equal loading of the samples. The co-staining of nuclei was performed using Hoechst 33342 (white). (A,B) Representative confocal images showing nuclear morphology and γH2AX staining in 14-3-3σ-knockout HCT116 (A) or wild-type HCT116 (B) cells after treatment for indicated time. (C) 14-3-3σ-knockout HCT116 cells were treated for 9 hours with doxorubicin after which the culture medium was replaced and cells were grown for an additional 24, 48 and 72 hours. Immunoblots of p53, phosphorylated ATM and γH2AX are shown. (D) Doxorubicin-induced DNA damage measured by alkaline comet assay. Cells were analysed by fluorescence microscopy and scored according to tail moment. Error bars represent the mean of three independent experiments±s.e.m. Scale bar: 20 μm. 1950

Long Term Aggresome Accumulation Leads to DNA Damage, p53-dependent Cell Cycle Arrest, and Steric Interference in Mitosis

“…As p21 regulates the cell cycle at the G 1 /S transition and induces G 1 phase arrest in response to a variety of stimuli, this is the likely cause of the reduced numbers of aggresome-containing cells entering mitosis. Any event in this pathway, from DSB formation, the activation of p53, and up-regulation of p21 to cell cycle arrest, can lead to caspase-dependent apoptosis (27,46,47). Driven by long term deposition of juxtanuclear aggresomes, these events may, therefore, contribute to the activation of apoptosis.…”

“…This leads to stabilization of p53, which thus promotes the transcription of cyclin-dependent kinase inhibitor p21 leading to arrest of the cell cycle at the G 1 /S transition (26) or which initiates apoptosis (27). We used immunoblotting to investigate whether P-p53 (Ser-15) is more prevalent in aggresome-containing cells and found that long term expression of EGFP-HDQ72 caused a gradual increase in P-p53 (Ser-15) levels over a 3-month period (Fig.…”

Hepatitis B virus pre‐S2 mutant large surface protein inhibits DNA double‐strand break repair and leads to genome instability in hepatocarcinogenesis

“…MNase-digestion assays, the standard method to detect DNA breaks in chromatin, showed that the pre-S 2 mutant LHBS + cells displayed increased DNA damage-induced degradation in the context of nucleosome structure, whereas the control and wild-type LHBS did not ( Figure 3E) [17]. Accumulating unrepaired DNA DSBs eventually leads to the formation of multinucleated cells, because of chromosome breakage-induced mitotic catastrophes [21]. Indeed, pre-S 2 mutant LHBS + and importin α1 knock-down cells, as well as the NBS1 knock-down cells, showed a significant number of multinucleated cells, which indicated abnormal chromosome breakages and cell-cycle control ( Figure 3F).…”

SETD2 reduction adversely affects the development of mouse early embryos

“…B, Quantification of the numbers of γ-H2AX foci per embryo in control and SETD2-KD group was analyzed (total number of embryos analyzed: n = 13 for control; n = 13 for SETD2-KD). 31 If the degree of damage is beyond the competence of recovery, cells never enter mitosis but undergo apoptotic cell senescence or death. *P < .05 vs control.…”