2026-06-14T06:44:08Zhttps://repisalud.isciii.es/rest/oai/requestoai:repisalud.isciii.es:20.500.12105/76732024-11-29T18:22:05Zcom_20.500.12105_2173com_20.500.12105_2051col_20.500.12105_19597
Repisalud
author
Díaz-Talavera, Alberto
author
Calvo, Patricia A
author
Gonzalez-Acosta, Daniel
author
Díaz, Marcos
author
Sastre-Moreno, Guillermo
author
Blanco-Franco, Luis
author
Guerra, Susana
author
Martínez-Jiménez, Maria I
author
Mendez, Juan
author
Blanco, Luis
funder
Unión Europea. Fondo Europeo de Desarrollo Regional (FEDER/ERDF)
funder
Ministerio de Economía y Competitividad (España)
funder
Fundación General (CSIC)
2019-05-24T08:41:08Z
2019-05-24T08:41:08Z
2019-02-04
Sci Rep. 2019 ;9(1):112.
10.1038/s41598-018-37439-0
2045-2322
2045-2322
Scientific reports
30718533
http://hdl.handle.net/20.500.12105/7673
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, Tyr100, as the main effector of sugar discrimination in human PrimPol and a change of Tyr100 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.
eng
DNA-POLYMERASE
ENZYME
RIBONUCLEOTIDE INCORPORATION
KINETIC-ANALYSIS
REPLICATION
PRIMASE
GENOME
RNA
NUCLEOTIDE
PROTEIN
A cancer-associated point mutation disables the steric gate of human PrimPol
journal article
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