Marteijn, J. A. et al. Nucleotide excision repair-induced H2A ubiquitination is dependent on MDC1 and RNF8 and reveals a universal DNA damage response. J. Cell Biol. 186, 835-847

Department of Genetics, Center for Biomedical Genetics, Erasmus Medical Center, 3015 GE Rotterdam, Netherlands.
The Journal of Cell Biology (Impact Factor: 9.69). 09/2009; 186(6):835-47. DOI: 10.1083/jcb.200902150
Source: PubMed

ABSTRACT Chromatin modifications are an important component of the of DNA damage response (DDR) network that safeguard genomic integrity. Recently, we demonstrated nucleotide excision repair (NER)-dependent histone H2A ubiquitination at sites of ultraviolet (UV)-induced DNA damage. In this study, we show a sustained H2A ubiquitination at damaged DNA, which requires dynamic ubiquitination by Ubc13 and RNF8. Depletion of these enzymes causes UV hypersensitivity without affecting NER, which is indicative of a function for Ubc13 and RNF8 in the downstream UV-DDR. RNF8 is targeted to damaged DNA through an interaction with the double-strand break (DSB)-DDR scaffold protein MDC1, establishing a novel function for MDC1. RNF8 is recruited to sites of UV damage in a cell cycle-independent fashion that requires NER-generated, single-stranded repair intermediates and ataxia telangiectasia-mutated and Rad3-related protein. Our results reveal a conserved pathway of DNA damage-induced H2A ubiquitination for both DSBs and UV lesions, including the recruitment of 53BP1 and Brca1. Although both lesions are processed by independent repair pathways and trigger signaling responses by distinct kinases, they eventually generate the same epigenetic mark, possibly functioning in DNA damage signal amplification.

  • Source
    • "It is conceivable that local or compartmentalized fluctuations of Ub levels might be achieved by bursts of DUB activity or, conversely , E3 ligase activity. In support of this possibility, FRAP experiments examining chromatin ubiquitylation at UV lesions indicated that Ub levels are highly dynamic at sites of DNA damage (Marteijn et al., 2009). "
    Dataset: OTUB1-2012
  • Source
    • "The two major DNA lesions induced by UV irradiation display different repair kinetics: while local induced 6-4PPs are repaired (depending on the dose) within ∼2 to 4 h; CPD repair is not achieved within 24 h [51]. Previous studies showed that accumulation of NER factors to LUD follow the repair kinetics of 6-4PP [27] [51]. It is unlikely that RPA accumulation to LUD at late time points is due to repair of 6-4PPs. "
    [Show abstract] [Hide abstract]
    ABSTRACT: The ability of replication protein A (RPA) to bind single-stranded DNA (ssDNA) underlines its crucial roles during DNA replication and repair. A combination of immunofluorescence and live cell imaging of GFP-tagged RPA70 revealed that RPA, in contrast to other replication factors, does not cluster into replication foci, which is explained by its short residence time at ssDNA. In addition to replication, RPA also plays a crucial role in both the pre- and post-incision steps of nucleotide excision repair (NER). Pre-incision factors like XPC and TFIIH accumulate rapidly at locally induced UV-damage and remain visible up to 4 h. However, RPA did not reach its maximum accumulation level until 3 h after DNA damage infliction and a chromatin-bound pool remained detectable up to 8 h, probably reflecting its role during the post-incision step of NER. During the pre-incision steps of NER, RPA could only be visualized at DNA lesions in incision deficient XP-F cells, however without a substantial increase in residence time at DNA damage. Together our data show that RPA is an intrinsically highly dynamic ssDNA-binding complex during both replication and distinct steps of NER.
    DNA Repair 12/2014; 24. DOI:10.1016/j.dnarep.2014.09.013 · 3.36 Impact Factor
  • Source
    • "This being so, how is DSB signaling mediated in the absence of H2AX phosphorylation in Arabidopsis? A possible answer comes from reports showing roles of modifications of other histones around DSB in mammals: ubiquitinylation of H2A by RNF8 is required for proper 53BP1 recruitment (Marteijn et al., 2009; Rossetto et al., 2010) and a role for histone lysine methylation in DSB repair is supported by the observation that H3K36me2 enhances DNA repair by NHEJ (Fnu et al., 2011). H3K36me2, once formed at DSB site, may create docking sites for other repair proteins, recruiting them for transcription and DNA repair. "
    [Show abstract] [Hide abstract]
    ABSTRACT: Failure to repair DNA double strand breaks (DSB) can lead to chromosomal rearrangements and eventually to cancer or cell death. Radiation and environmental pollutants induce DSB and this is of particular relevance to plants due to their sessile life style. DSB also occur naturally in cells during DNA replication and programmed induction of DSB initiates the meiotic recombination essential for gametogenesis in most eukaryotes. The linear nature of most eukaryotic chromosomes means that each chromosome has two "broken" ends. Chromosome ends, or telomeres, are protected by nucleoprotein caps which avoid their recognition as DSB by the cellular DNA repair machinery. Deprotected telomeres are recognized as DSB and become substrates for recombination leading to chromosome fusions, the "bridge-breakage-fusion" cycle, genome rearrangements and cell death. The importance of repair of DSB and the severity of the consequences of their misrepair have led to the presence of multiple, robust mechanisms for their detection and repair. After a brief overview of DSB repair pathways to set the context, we present here an update of current understanding of the detection and signaling of DSB in the plant, Arabidopsis thaliana.
    Frontiers in Plant Science 10/2013; 4:405. DOI:10.3389/fpls.2013.00405 · 3.95 Impact Factor
Show more