Polμ tumor variants decrease the efficiency and accuracy of NHEJ
| Autor: | Alberto Díaz-Talavera, Luis Blanco, John M. Pryor, José F. Ruiz, Dale A. Ramsden, Guillermo Sastre-Moreno |
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| Přispěvatelé: | European Commission, Consejo Superior de Investigaciones Científicas (España), American Cancer Society, Ministerio de Economía y Competitividad (España), Universidad de Sevilla. Departamento de Bioquímica Vegetal y Biología Molecular, Ministerio de Economía y Competitividad (MINECO). España |
| Rok vydání: | 2017 |
| Předmět: |
Models
Molecular 0301 basic medicine DNA End-Joining Repair Protein Conformation DNA polymerase Protein domain Glycine Mutation Missense Electrophoretic Mobility Shift Assay Sequence alignment DNA-Directed DNA Polymerase Computational biology Genome Integrity Repair and Replication Arginine medicine.disease_cause Conserved sequence 03 medical and health sciences Protein Domains Genetics medicine Humans Point Mutation Conserved Sequence Mutation Sequence Homology Amino Acid biology Point mutation fungi Mutagenesis Neoplasm Proteins enzymes and coenzymes (carbohydrates) 030104 developmental biology Oligodeoxyribonucleotides biology.protein Sequence Alignment |
| Zdroj: | Nucleic Acids Research Digital.CSIC. Repositorio Institucional del CSIC instname idUS. Depósito de Investigación de la Universidad de Sevilla |
| ISSN: | 1362-4962 0305-1048 |
| DOI: | 10.1093/nar/gkx625 |
| Popis: | The non homologous end-joining (NHEJ) pathway of double-strand break (DSB) repair often requires DNA synthesis to fill the gaps generated upon alignment of the broken ends, a complex task performed in human cells by two specialized DNA polymerases, Polλ and Polμ. It is now well established that Polμ is the one adapted to repair DSBs with non-complementary ends, the most challenging scenario, although the structural basis and physiological implications of this adaptation are not fully understood. Here, we demonstrate that two human Polμ point mutations, G174S and R175H, previously identified in two different tumor samples and affecting two adjacent residues, limit the efficiency of accurate NHEJ by Polμ in vitro and in vivo. Moreover, we show that this limitation is the consequence of a decreased template dependency during NHEJ, which renders the error-rate of the mutants higher due to the ability of Polμ to randomly incorporate nucleotides at DSBs. These results highlight the relevance of the 8 kDa domain of Polμ for accurate and efficient NHEJ, but also its contribution to the error-prone behavior of Polμ at 2-nt gaps. This work provides the first demonstration thatmutations affecting Polμ identified in tumors can alter the efficiency and fidelity of NHEJ. Spanish Ministry of Economy and Competitiveness [BFU2012-37969, BFU2015-65880-P to L.B., BFU2013-44343-P to J.F.R.]; European Commission (European Regional Development Fund [RYC-2011-08752 to J.F.R.]; JAE-predoctoral fellowship from the Spanish Research Council (CSIC) (to G.S.-M.); Ramsden laboratory was supported by a postdoctoral fellowship from the American Cancer Society [PF-14-0438-01-DMC to J.M.P.]; NCI grant [CA097096 to D.A.R.]. Funding for open access charge: Spanish Ministry of Economy and Competitiveness [BFU2012-37969, BFU2015-65880-P to L.B.]. |
| Databáze: | OpenAIRE |
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