2026-06-14T03:54:51Zhttps://repisalud.isciii.es/rest/oai/requestoai:repisalud.isciii.es:20.500.12105/137182024-11-29T15:42:40Zcom_20.500.12105_2173com_20.500.12105_2051col_20.500.12105_19597
Repisalud
author
Calvo, Patricia A
author
Martínez-Jiménez, María I
author
Díaz, Marcos
author
Stojkovic, Gorazd
author
Kasho, Kazutoshi
author
Guerra, Susana
author
Wanrooij, Sjoerd
author
Blanco, Luis
author
Mendez, Juan
funder
Unión Europea. Fondo Europeo de Desarrollo Regional (FEDER/ERDF)
funder
Knut and Alice Wallenberg Foundation
funder
Ministerio de Economía, Industria y Competitividad (España)
2022-03-03T12:07:56Z
2022-03-03T12:07:56Z
2021-08-20
Nucleic Acids Res . 2021;49(14):8199-8213.
10.1093/nar/gkab634
1362-4962
Nucleic acids research
34302490
http://hdl.handle.net/20.500.12105/13718
PrimPol is the second primase in human cells, the first with the ability to start DNA chains with dNTPs. PrimPol contributes to DNA damage tolerance by restarting DNA synthesis beyond stalling lesions, acting as a TLS primase. Multiple alignment of eukaryotic PrimPols allowed us to identify a highly conserved motif, WxxY near the invariant motif A, which contains two active site metal ligands in all members of the archeo-eukaryotic primase (AEP) superfamily. In vivo and in vitro analysis of single variants of the WFYY motif of human PrimPol demonstrated that the invariant Trp87 and Tyr90 residues are essential for both primase and polymerase activities, mainly due to their crucial role in binding incoming nucleotides. Accordingly, the human variant F88L, altering the WFYY motif, displayed reduced binding of incoming nucleotides, affecting its primase/polymerase activities especially during TLS reactions on UV-damaged DNA. Conversely, the Y89D mutation initially associated with High Myopia did not affect the ability to rescue stalled replication forks in human cells. Collectively, our data suggest that the WFYY motif has a fundamental role in stabilizing the incoming 3'-nucleotide, an essential requisite for both its primase and TLS abilities during replication fork restart.
eng
DNA-POLYMERASE-GAMMA
PRIMASE
DOMAIN
ARCHITECTURE
INSIGHTS
REVEALS
RNA
Motif WFYY of human PrimPol is crucial to stabilize the incoming 3'-nucleotide during replication fork restart.
journal article
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