Coordination of Primer Initiation Within the Catalytic Domain of Human PrimPol.
J Mol Biol
; 435(24): 168338, 2023 12 15.
Article
in En
| MEDLINE
| ID: mdl-37923120
ABSTRACT
To facilitate the eukaryotic repriming pathway of DNA damage tolerance, PrimPol synthesises de novo oligonucleotide primers downstream of polymerase-stalling obstacles. These primers enable replicative polymerases to resume synthesis and ensure the timely completion of DNA replication. Initiating synthesis de novo requires the coordination of single-stranded DNA, initiating nucleotides, and metal ions within PrimPol's active site to catalyze the formation of the first phosphodiester bond. Here we examine the interactions between human PrimPol's catalytic domain, nucleotides, and DNA template during each of the various catalytic steps to determine the 'choreography' of primer synthesis, where substrates bind in an ordered manner. Our findings show that the ability of PrimPol to conduct de novo primer synthesis is underpinned by a network of stabilising interactions between the enzyme, template, and nucleotides, as we previously observed for related primase CRISPR-Associated Prim-Pol (CAPP). Together, these findings establish a detailed model for the initiation of DNA synthesis by human PrimPol, which appears highly conserved.
Key words
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Catalytic Domain
/
DNA-Directed DNA Polymerase
/
DNA Replication
Limits:
Humans
Language:
En
Journal:
J Mol Biol
Year:
2023
Document type:
Article
Affiliation country:
Reino Unido