SUMO-targeted ubiquitin E3 ligase RNF4 is required for the response of human cells to DNA damage
| Autor: | Joy S. Chua, Anne Seifert, Ronald T. Hay, Filip Golebiowski, Jean-François Maure, Yili Yin |
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| Rok vydání: | 2012 |
| Předmět: |
DNA Repair
DNA damage DNA repair Ubiquitin-Protein Ligases SUMO protein Cell Cycle Proteins DDB1 Cell Line Tumor Genetics Animals Humans DNA Breaks Double-Stranded Homologous Recombination Replication protein A Adaptor Proteins Signal Transducing biology Ubiquitin Nuclear Proteins G2-M DNA damage checkpoint Molecular biology Ubiquitin ligase MDC1 Rats biology.protein Trans-Activators Developmental Biology DNA Damage HeLa Cells Transcription Factors Research Paper |
| Zdroj: | Genesdevelopment. 26(11) |
| ISSN: | 1549-5477 |
| Popis: | Here we demonstrate that RNF4, a highly conserved small ubiquitin-like modifier (SUMO)-targeted ubiquitin E3 ligase, plays a critical role in the response of mammalian cells to DNA damage. Human cells in which RNF4 expression was ablated by siRNA or chicken DT40 cells with a homozygous deletion of the RNF4 gene displayed increased sensitivity to DNA-damaging agents. Recruitment of RNF4 to double-strand breaks required its RING and SUMO interaction motif (SIM) domains and DNA damage factors such as NBS1, mediator of DNA damage checkpoint 1 (MDC1), RNF8, 53BP1, and BRCA1. In the absence of RNF4, these factors were still recruited to sites of DNA damage, but 53BP1, RNF8, and RNF168 displayed delayed clearance from such foci. SILAC-based proteomics of SUMO substrates revealed that MDC1 was SUMO-modified in response to ionizing radiation. As a consequence of SUMO modification, MDC1 recruited RNF4, which mediated ubiquitylation at the DNA damage site. Failure to recruit RNF4 resulted in defective loading of replication protein A (RPA) and Rad51 onto ssDNA. This appeared to be a consequence of reduced recruitment of the CtIP nuclease, resulting in inefficient end resection. Thus, RNF4 is a novel DNA damage-responsive protein that plays a role in homologous recombination and integrates SUMO modification and ubiquitin signaling in the cellular response to genotoxic stress. |
| Databáze: | OpenAIRE |
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