In vivo genotoxicity of ortho-phenylphenol, biphenyl, and thiabendazole detected in multiple mouse organs by the alkaline single cell gel electrophoresis assay

Norberg

Journal of Cell Science

et al. 2011

) relating to the article 'Chromosomal breaks during mitotic catastrophe trigger γH2AX-ATM-p53-mediated apoptosis' by Gabriela Imreh, Helin Vakifahmetoglu Norberg, Stefan Imreh and Boris Zhivotovsky. J. Cell Sci. 2011Sci. 124, 2951Sci. -2963Sci. (doi: 10.1242).Concerns were raised regarding some of the data in Fig. 1A of the above-named paper. An investigation carried out by Karolinska Institutet concluded that no wrongdoing had occurred, and that no further action was required. An in-house review of the original data by Journal of Cell Science reached a similar conclusion.Journal of Cell Science is publishing this Note to alert readers that the concerns raised about this paper have been resolved. This course of action follows the advice set out by COPE (Committee on Publication Ethics), of which Journal of Cell Science is a member. Concerns have been raised regarding some of the data in Fig. 1A of the above-named paper. Journal of Cell Science is publishing this note to make readers aware of this issue, and we will provide further information once the issue has been resolved. 3419This course of action follows the advice set out by COPE (Committee on Publication Ethics), of which Journal of Cell Science is a member. There was an error published in J. Cell Sci. 124, 2951-2963. 1979In 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. Science (2016Science ( ) 129, 1950Science ( doi:10.1242 © 2016. Published by The Company of Biologists Ltd | Journal of Cell Journal of Cell ScienceChromosomal breaks during mitotic catastrophe trigger cH2AX-ATM-p53-mediated apoptosis Summary Although the cause...

“…The comet assay was performed using the method described earlier with some modifications (Sasaki et al, 1997). Briefly, cells were exposed to 600 nM doxorubicin for 9, 16 and 24 hours.…”

Section: Comet Assaymentioning

confidence: 99%

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

Norberg

Journal of Cell Science

et al. 2011

“…Sin embargo, informaron sobre su capacidad mutagénica cuando es fotoactivado con luz UV. El ensayo cometa se ha utilizado en estudios in vivo con ratones ddY y CD-1, a los que se les suministraron dosis orales de tiabendazol (200-2000 mg/kg) para determinar el daño al ADN en varios ór-ganos como el estómago, hígado, pulmón, entre otros, y se encontraron incrementos significativos en la longitud del cometa (Sasaki et al 1997(Sasaki et al , 2002. Sin embargo, la técnica no ha sido empleada para evaluar el efecto genotóxico del tiabendazol en células humanas.…”

Section: Discussionunclassified

Genotoxicidad de tres plaguicidas utilizados en la actividad bananera de Costa Rica

Vindas

Ortíz

Ramírez

et al. 2014

RBT

Genotoxicity of three pesticides used in Costa Rican banana plantations. The in vitro genotoxicity of imazalil and thiabendazole fungicides and the insecticide chlorpyrifos, compounds used in Costa Rican banana plantations, was evaluated with the single-cell gel electrophoresis technique (comet assay). The comet assay is a simple, rapid and low cost technique for quantification of DNA damage. This assay detects DNA single-strand breaks and alkali-labile sites in individual cells. The effects were analyzed by using human lymphocytes exposed to doses of 0, 25, 50, 75 and 100 µg/ml of each pesticide for 30 min at 37°C. The cells were embedded in agarose, lysed, subjected to alkaline electrophoresis (pH> 13) for 20 min at 25V, neutralized and dehydrated to be stained with a fluorescent dye and later comets visualization with the epifluorescence microscope. Chlorpyrifos and imazalil induced significant DNA damage in a dose-dependent manner. Chlorpyrifos was the major inductor of DNA breaks. These results indicate that both are genotoxic compounds in vitro. Thiabendazole fungicide did not induced DNA damage using the comet assay for all concentrations tested. Rev. Biol. Trop. 52(3): 601-609. Epub 2004 Dic 15.

“…The alkaline comet assay was performed using the method described previously with some modifications (Sasaki et al 1997). Briefly, cells were transfected with siRNA as described above for 24 h and irradiated.…”

Section: Comet Assaymentioning

confidence: 99%

The scaffold protein WRAP53β orchestrates the ubiquitin response critical for DNA double-strand break repair

et al. 2014

The WD40 domain-containing protein WRAP53b (WD40 encoding RNA antisense to p53; also referred to as WDR79/TCAB1) controls trafficking of splicing factors and the telomerase enzyme to Cajal bodies, and its functional loss has been linked to carcinogenesis, premature aging, and neurodegeneration. Here, we identify WRAP53b as an essential regulator of DNA double-strand break (DSB) repair. WRAP53b rapidly localizes to DSBs in an ATM-, H2AX-, and MDC1-dependent manner. We show that WRAP53b targets the E3 ligase RNF8 to DNA lesions by facilitating the interaction between RNF8 and its upstream partner, MDC1, in response to DNA damage. Simultaneous binding of MDC1 and RNF8 to the highly conserved WD40 scaffold domain of WRAP53b facilitates their interaction and accumulation of RNF8 at DSBs. In this manner, WRAP53b controls proper ubiquitylation at DNA damage sites and the downstream assembly of 53BP1, BRCA1, and RAD51. Furthermore, we reveal that knockdown of WRAP53b impairs DSB repair by both homologous recombination (HR) and nonhomologous end-joining (NHEJ), causes accumulation of spontaneous DNA breaks, and delays recovery from radiation-induced cell cycle arrest. Our findings establish WRAP53b as a novel regulator of DSB repair by providing a scaffold for DNA repair factors.