The rapid recruitment of PNKP to damage sites (red) (Segal-Raz et al., 2011) marks their location. Cells were depleted of endogenous UBE4A using RNAi and co-transfected with vectors expressing siRNA-resistant cDNAs of GFP-UBE4A or GFP-UBE4AΔU-box, together with DsRed2-tagged polynucleotide kinase-phosphatase (PNKP). (A) Dynamics of UBE4A relocalization to damage sites in live cells. Our data uncover a critical regulatory level in the DSB response and underscore the importance of fine-tuning the complex DDR network for accurate and balanced execution of DSB repair.ĭNA damage UBE4A double-strand breaks genome stability ubiquitin.Ĭopyright © 2018 Elsevier Inc. This pathway is essential for optimal end resection at DSBs, and its abrogation leads to upregulation of the highly mutagenic alternative end-joining repair at the expense of error-free homologous recombination repair. This step is required for timely recruitment of the RAP80 and BRCA1 proteins and proper organization of RAP80- and BRCA1-associated protein complexes at DSB sites. UBE4A's recruitment to sites of DNA damage is dependent on primary E3 ligases in the DDR and promotes enhancement and sustainment of K48- and K63-linked ubiquitin chains at these sites. We identified a critical player in DDR fine-tuning: the E3/E4 ubiquitin ligase UBE4A. Electronic address: breaks (DSBs) are critical DNA lesions that robustly activate the elaborate DNA damage response (DDR) network. 9 The David and Inez Myers Laboratory for Cancer Research, Department of Human Molecular Genetics and Biochemistry, Sackler School of Medicine, Tel Aviv University, Tel Aviv, Israel.8 State Key Laboratory of Stem Cell and Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, China.7 Department of Biochemistry and Protein Network Research Center, Yonsei University, 134 Shinchon-Dong, Seodaemoon-Gu, Seoul, Korea.6 Departments of Pathology and Biochemistry and Molecular Biology, Dalhousie University, Halifax, NS, Canada.5 National Cancer Institute, NIH, Bethesda, MD, USA.4 Centro Andaluz de Biología Molecular y Medicina Regenerativa (CABIMER) and Department of Genetics, University of Sevilla, Sevilla, Spain.3 Perlmutter NYU Cancer Center and Department of Biochemistry and Molecular Pharmacology, New York University School of Medicine, New York, NY, USA.2 Cancer Research UK and Medical Research Council Oxford Institute for Radiation Oncology, Department of Oncology, University of Oxford, Oxford, UK.1 The David and Inez Myers Laboratory for Cancer Research, Department of Human Molecular Genetics and Biochemistry, Sackler School of Medicine, Tel Aviv University, Tel Aviv, Israel.
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