A cancer-associated point mutation disables the steric gate of human PrimPol
| Autor: | Patricia A. Calvo, Daniel González-Acosta, Marcos Díaz, Juan Méndez, Alberto Díaz-Talavera, María I. Martínez-Jiménez, Susana Guerra, Guillermo Sastre-Moreno, Luis Blanco, Luis Blanco-Franco |
|---|---|
| Přispěvatelé: | Ministerio de Economía y Competitividad (España), European Commission, Centro Nacional de Investigaciones Oncológicas (España), Unión Europea. Fondo Europeo de Desarrollo Regional (FEDER/ERDF), Fundación General (CSIC) |
| Jazyk: | angličtina |
| Rok vydání: | 2019 |
| Předmět: |
0301 basic medicine
Models Molecular DNA polymerase DNA-POLYMERASE RIBONUCLEOTIDE INCORPORATION Drug Resistance PROTEIN lcsh:Medicine DNA-Directed DNA Polymerase medicine.disease_cause Mice 0302 clinical medicine Neoplasms Hydroxyurea Nucleotide lcsh:Science Polymerase PRIMASE chemistry.chemical_classification Mutation Multidisciplinary biology Chemistry Nucleotides Cell Cycle Cell biology GENOME Primase ENZYME NUCLEOTIDE Pyrimidine dimer DNA Primase Article Cell Line 03 medical and health sciences medicine Animals Humans Point Mutation Computer Simulation Histidine Point mutation KINETIC-ANALYSIS lcsh:R RNA Multifunctional Enzymes 030104 developmental biology REPLICATION biology.protein Tyrosine lcsh:Q 030217 neurology & neurosurgery |
| Zdroj: | Scientific Reports Scientific Reports, Vol 9, Iss 1, Pp 1-13 (2019) Digital.CSIC. Repositorio Institucional del CSIC instname Repisalud Instituto de Salud Carlos III (ISCIII) |
| ISSN: | 2045-2322 |
| Popis: | PrimPol is a human primase/polymerase specialized in re-starting stalled forks by repriming beyond lesions such as pyrimidine dimers, and replication-perturbing structures including G-quadruplexes and R-loops. Unlike most conventional primases, PrimPol proficiently discriminates against ribonucleotides (NTPs), being able to start synthesis using deoxynucleotides (dNTPs), yet the structural basis and physiological implications for this discrimination are not understood. In silico analyses based on the three-dimensional structure of human PrimPol and related enzymes enabled us to predict a single residue, Tyr, as the main effector of sugar discrimination in human PrimPol and a change of Tyr to histidine to boost the efficiency of NTP incorporation. We show here that the Y100H mutation profoundly stimulates NTP incorporation by human PrimPol, with an efficiency similar to that for dNTP incorporation during both primase and polymerase reactions in vitro. As expected from the higher cellular concentration of NTPs relative to dNTPs, Y100H expression in mouse embryonic fibroblasts and U2OS osteosarcoma cells caused enhanced resistance to hydroxyurea, which decreases the dNTP pool levels in S-phase. Remarkably, the Y100H PrimPol mutation has been identified in cancer, suggesting that this mutation could be selected to promote survival at early stages of tumorigenesis, which is characterized by depleted dNTP pools. Spanish Ministry of Economy and Competitiveness (MINECO; BFU2012–3769, BFU2014–51672-REDC and BFU2015–65880-P (co-funded with European Union FEDER funds) to L.B.; BFU2013–49153-P and BFU2016–80402-R (co-funded with European Union FEDER funds) to J.M.). A.D.T., P.A.C. and D.G.A. are recipients of MINECO FPI-predoctoral fellowships. M.D. was the recipient of a FPI-predoctoral fellowship from Programa de Excelencia “Severo Ochoa” (CNIO-MINECO) |
| Databáze: | OpenAIRE |
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