The DNA ligands Arg47 and Arg76 are crucial for catalysis by human PrimPol.

Autor: Boldinova EO; National Research Center «Kurchatov Institute» - Institute of Molecular Genetics, Moscow, 123182, Russia., Manukyan АА; National Research Center «Kurchatov Institute» - Institute of Molecular Genetics, Moscow, 123182, Russia; D. Mendeleev University of Chemical Technology of Russia (D. Mendeleyev University, MUCTR), Moscow, 125047, Russia., Makarova АV; National Research Center «Kurchatov Institute» - Institute of Molecular Genetics, Moscow, 123182, Russia. Electronic address: amakarova-img@yandex.ru.
Jazyk: angličtina
Zdroj: DNA repair [DNA Repair (Amst)] 2021 Apr; Vol. 100, pp. 103048. Date of Electronic Publication: 2021 Jan 29.
DOI: 10.1016/j.dnarep.2021.103048
Abstrakt: Human primase and DNA polymerase PrimPol re-starts stalled replication forks by repriming downstream DNA lesions and protects cells against DNA damage. Structure of the catalytic core of PrimPol with DNA primer, template and incoming dATP was solved but the mechanisms of DNA polymerase and primase activities of PrimPol are not fully understood. In this work, using site-directed mutagenesis we biochemically analyzed the role of active site residues Arg47 and Arg76 contacting DNA template in DNA polymerase and primase activities of PrimPol. The substitution R47A diminished the DNA polymerase and primase activities of PrimPol whereas the single amino acid substitution R76A caused almost complete loss of catalytic activities. Both amino acid substitutions affected the spectrum of dNMPs incorporation on undamaged DNA templates and opposite 8-oxoguanine. Finally, substitutions of the Arg47 and Arg76 residues attenuated the formation of the stable PrimPol:DNA complex in the presence of ATP/dNTPs. Together, these findings suggest a key role of the Arg47 and Arg76 in DNA synthesis by PrimPol.
(Copyright © 2021 Elsevier B.V. All rights reserved.)
Databáze: MEDLINE
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