Nucleotide excision repair–induced H2A ubiquitination is dependent on MDC1 and RNF8 and reveals a universal DNA damage response

Marteijn, Jürgen A.; Bekker‐Jensen, Simon; Mailand, Niels; Lans, Hannes; Schwertman, Petra; Gourdin, Audrey M.; Dantuma, Nico P.; Lukáš, Jiří; Vermeulen, Wim

doi:10.1083/jcb.200902150

Cited by 170 publications

(187 citation statements)

References 61 publications

(109 reference statements)

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“…Moreover, Jeggo and co-workers (29) reported that replication-independent ATR signaling requires NBS1, 53BP1, and MDC1, but not H2AX, to induce G 2 /M arrest. Recently, MDC1 was reported to accumulate at locally UV-damaged subnuclear regions in an NER-and ATR-dependent manner, which recruits RNF8 ubiquitinating histone H2A and hereby 53BP1 and BRCA1 (30). In our experiments, however, local accumulation of MRE11 after micropore UV irradiation was markedly diminished by KU-55933 (Fig.…”

Section: Discussioncontrasting

confidence: 52%

Nucleotide Excision Repair-dependent DNA Double-strand Break Formation and ATM Signaling Activation in Mammalian Quiescent Cells

Wakasugi

Sasaki

Matsumoto

et al. 2014

Journal of Biological Chemistry

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Background:In quiescent human cells, UV induces histone H2AX phosphorylation by ATR in a nucleotide excision repair (NER)-dependent manner. Results: UV also activates ATM in response to NER-mediated DNA double-strand break (DSB). Conclusion:The NER reaction in quiescent cells potentially generates multiple types of secondary DNA damage. Significance: This work highlights the importance of our understanding of the DNA damage response in quiescent cells.

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Section: Discussioncontrasting

confidence: 52%

Nucleotide Excision Repair-dependent DNA Double-strand Break Formation and ATM Signaling Activation in Mammalian Quiescent Cells

Wakasugi

Sasaki

Matsumoto

et al. 2014

Journal of Biological Chemistry

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“…While we were unable to detect phosphorylation of histone H2AX with 30 J/m 2 of UVC in our cells without BrdU-labelling, NER-coupled phosphorylation has been reported in the G1 or G2 phase with 60 to 100 J/m 2 of UVC (32,33). In quiescent cells, phosphorylation was detected after 10 J/m 2 or less (33,34).…”

Section: Experimental Designcontrasting

confidence: 71%

“…In quiescent cells, phosphorylation was detected after 10 J/m 2 or less (33,34). Thus, the UVC dose that dose not elicit any significant confounding effects, needs to be determined.…”

Section: Experimental Designmentioning

confidence: 99%

Creating localized DNA double-strand breaks with microirradiation

et al. 2011

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SUMMARYWe describe a protocol for creating localized DNA double-strand breaks (DSBs) with minimal requirements that can be applied in cell biology and molecular biology. This protocol is based on the combination of 5-bromo-2'-deoxyuridine (BrdU)-labelling and ultraviolet C (UVC)-irradiation through porous membranes. Cells are labelled with 10 µM BrdU for 48-72 hours, washed with Ca 2+ and Mg 2+ -free phosphate buffered saline (PBS (-)), covered by polycarbonate membranes with micro-pores, and exposed to UVC-light. Using this protocol, localized DSBs are created within subnuclear areas, irrespective of the cell cycle phase.Recruitment of proteins involved in DNA repair, DNA damage response, chromatin remodelling and histone modifications can be visualized without any specific equipment. The quality is the same as that obtained by the laser micro-irradiation or any other focal irradiation.DSBs become visible within 30 minutes of UVC irradiation.

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“…Upon binding to gH2AX, the MDC1 protein mediates signal expansion on the DNA molecule (Stucki et al 2005) and serves as an anchor for subsequent recruitment of the E3 ubiquitin ligases RNF8 and RNF168. RNF8 is the first E3 ligase to be recruited to DSBs, which, in conjunction with RNF168, ubiquitylates flanking chromatin (including histones H2AX and H2A) and promotes efficient assembly of the downstream repair factors 53BP1, BRCA1, and RAD51 at the site of damage (Huen et al 2007;Mailand et al 2007;Marteijn et al 2009;Meerang et al 2011). The DDR involves accumulation of DNA repair factors at DNA lesions, a process mediated largely by protein interactions.…”

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confidence: 99%

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

et al. 2014

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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.

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Nucleotide excision repair–induced H2A ubiquitination is dependent on MDC1 and RNF8 and reveals a universal DNA damage response

Abstract: The epigenetic mark indicative of DNA UV damage or double-strand breaks is achieved via a common pathway regardless of the cause of damage.

Cited by 170 publications

References 61 publications

Nucleotide Excision Repair-dependent DNA Double-strand Break Formation and ATM Signaling Activation in Mammalian Quiescent Cells

Nucleotide Excision Repair-dependent DNA Double-strand Break Formation and ATM Signaling Activation in Mammalian Quiescent Cells

Creating localized DNA double-strand breaks with microirradiation

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

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