A key interaction with RPA orients XPA in NER complexes
| Autor: | Michal Hammel, Norie Sugitani, Daniel J. Rosenberg, Kateryna V Le Meur, Agnieszka M. Topolska-Woś, Orlando D. Schärer, Remy Le Meur, Hyunsuk Kim, Jung-Eun Yeo, Walter J. Chazin, John J Cordoba |
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| Přispěvatelé: | Lawrence Berkeley National Laboratory [Berkeley] (LBNL) |
| Rok vydání: | 2020 |
| Předmět: |
Models
Molecular endocrine system Magnetic Resonance Spectroscopy DNA Repair DNA Single-Stranded Biology 03 medical and health sciences chemistry.chemical_compound 0302 clinical medicine Dna genetics Functional importance Structural Biology Single-Stranded Models Replication Protein A Information and Computing Sciences Genetics Humans ComputingMilieux_MISCELLANEOUS 030304 developmental biology 0303 health sciences Extramural Binding protein Molecular DNA Biological Sciences Cell biology [SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry Molecular Biology/Biomolecules [q-bio.BM] Xeroderma Pigmentosum Group A Protein [SDV.BBM.BP]Life Sciences [q-bio]/Biochemistry Molecular Biology/Biophysics DNA-Binding Proteins [SDV.BBM.BS]Life Sciences [q-bio]/Biochemistry Molecular Biology/Biomolecules [q-bio.BM] enzymes and coenzymes (carbohydrates) chemistry Docking (molecular) 030220 oncology & carcinogenesis Xpa gene Environmental Sciences Nucleotide excision repair DNA Damage Protein Binding Developmental Biology |
| Zdroj: | Nucleic acids research, vol 48, iss 4 Nucleic Acids Research Nucleic Acids Research, Oxford University Press, 2020, 48 (4), pp.2173-2188. ⟨10.1093/nar/gkz1231⟩ |
| ISSN: | 0305-1048 1362-4962 |
| DOI: | 10.1093/nar/gkz1231⟩ |
| Popis: | The XPA protein functions together with the single-stranded DNA (ssDNA) binding protein RPA as the central scaffold to ensure proper positioning of repair factors in multi-protein nucleotide excision repair (NER) machinery. We previously determined the structure of a short motif in the disordered XPA N-terminus bound to the RPA32C domain. However, a second contact between the XPA DNA-binding domain (XPA DBD) and the RPA70AB tandem ssDNA-binding domains, which is likely to influence the orientation of XPA and RPA on the damaged DNA substrate, remains poorly characterized. NMR was used to map the binding interfaces of XPA DBD and RPA70AB. Combining NMR and X-ray scattering data with comprehensive docking and refinement revealed how XPA DBD and RPA70AB orient on model NER DNA substrates. The structural model enabled design of XPA mutations that inhibit the interaction with RPA70AB. These mutations decreased activity in cell-based NER assays, demonstrating the functional importance of XPA DBD–RPA70AB interaction. Our results inform ongoing controversy about where XPA is bound within the NER bubble, provide structural insights into the molecular basis for malfunction of disease-associated XPA missense mutations, and contribute to understanding of the structure and mechanical action of the NER machinery. |
| Databáze: | OpenAIRE |
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