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: 9.83). 09/2009; 186(6):835-47. DOI: 10.1083/jcb.200902150
Source: PubMed


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.

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    • "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). "
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    • "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. "
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    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 · 3.11 Impact Factor
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    • "Studies over the last few years in the model plant Arabidopsis have greatly advanced our knowledge about the roles of H2Aub1 and H2Bub1 in transcription regulation in plant growth and development. In view of additional functions described in animal cells for both H2Aub1 and H2Bub1 in DNA damage repair (Bergink et al., 2006; Marteijn et al., 2009; Chernikova et al., 2010; Ginjala et al., 2011; Moyal et al., 2011; Nakamura et al., 2011), it is anticipated that more roles of H2Aub1 and H2Bub1 in plant response to environmental stresses are waiting to be uncovered. Mutagenesis of enzymes involved in H2Aub1 and H2Bub1 deposition or removal is required to address the question whether these enzymes effectively exert their biological functions via H2Aub1 and H2Bub1. "
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    ABSTRACT: Polyubiquitin chain deposition on a target protein frequently leads to proteasome-mediated degradation whereas monoubiquitination modifies target protein property and function independent of proteolysis. Histone monoubiquitination occurs in chromatin and is in nowadays recognized as one critical type of epigenetic marks in eukaryotes. While H2A monoubiquitination (H2Aub1) is generally associated with transcription repression mediated by the Polycomb pathway, H2Bub1 is involved in transcription activation. H2Aub1 and H2Bub1 levels are dynamically regulated via deposition and removal by specific enzymes. We review knows and unknowns of dynamic regulation of H2Aub1 and H2Bub1 deposition and removal in plants and highlight the underlying crucial functions in gene transcription, cell proliferation/differentiation, and plant growth and development. We also discuss crosstalks existing between H2Aub1 or H2Bub1 and different histone methylations for an ample mechanistic understanding.
    Frontiers in Plant Science 03/2014; 5:83. DOI:10.3389/fpls.2014.00083 · 3.95 Impact Factor
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