XLF acts as a flexible connector during non-homologous end joining
| Autor: | Felicia Wednesday Lopezcolorado, Joseph J. Loparo, Sean M. Carney, Metztli Cisneros-Aguirre, Sadie C. Piatt, Jeremy M. Stark, Andrew T. Moreno |
|---|---|
| Rok vydání: | 2020 |
| Předmět: |
DNA End-Joining Repair
QH301-705.5 DNA repair Structural Biology and Molecular Biophysics Xenopus Science Immunoblotting Complex formation Xenopus Proteins single-molecule FRET non-homologous end joining General Biochemistry Genetics and Molecular Biology Xenopus laevis Cable gland DNA double strand break chemistry.chemical_compound Animals Biology (General) Ovum Double strand Physics General Immunology and Microbiology biology Chemistry General Neuroscience Synapsis General Medicine Single-molecule FRET DNA repair protein XRCC4 Chromosomes and Gene Expression biology.organism_classification Cell biology DNA-Binding Proteins Non-homologous end joining DNA Repair Enzymes Förster resonance energy transfer Structural biology Chromatography Gel Biophysics Medicine Sequence Alignment DNA Research Article |
| Zdroj: | eLife, Vol 9 (2020) eLife |
| ISSN: | 2050-084X |
| DOI: | 10.7554/elife.61920 |
| Popis: | Non-homologous end joining (NHEJ) is the predominant pathway that repairs DNA double-strand breaks in vertebrates. During NHEJ DNA ends are held together by a multi-protein synaptic complex until they are ligated. Here, we use Xenopus laevis egg extract to investigate the role of the intrinsically disordered C-terminal tail of the XRCC4-like factor (XLF), a critical factor in end synapsis. We demonstrate that the XLF tail along with the Ku-binding motif (KBM) at the extreme C-terminus are required for end joining. Although the underlying sequence of the tail can be varied, a minimal tail length is required for NHEJ. Single-molecule FRET experiments that observe end synapsis in real-time show that this defect is due to a failure to closely align DNA ends. Our data supports a model in which a single C-terminal tail tethers XLF to Ku, while allowing XLF to form interactions with XRCC4 that enable synaptic complex formation. |
| Databáze: | OpenAIRE |
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