Communication between DNA polymerases and Replication Protein A within the archaeal replisome

Markel Martínez-Carranza, Léa Vialle, Clément Madru, Florence Cordier, Ayten Dizkirici Tekpinar, Ahmed Haouz, Pierre Legrand, Rémy A. Meur, Patrick England, Rémi Dulermo, J. Iñaki Guijarro, Ghislaine Henneke () and Ludovic Sauguet ()
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Markel Martínez-Carranza: Université Paris Cité, CNRS UMR 3528
Léa Vialle: Univ Brest, Ifremer, CNRS, Biologie et Ecologie des Ecoystèmes marins profonds (BEEP)
Clément Madru: Université Paris Cité, CNRS UMR 3528
Florence Cordier: Université Paris Cité, CNRS UMR 3528
Ayten Dizkirici Tekpinar: Université Paris Cité, CNRS UMR 3528
Ahmed Haouz: Université Paris Cité, CNRS UMR 3528
Pierre Legrand: L’Orme des Merisiers
Rémy A. Meur: Université Paris Cité, CNRS UMR 3528
Patrick England: Université Paris Cité, CNRS UMR 3528
Rémi Dulermo: Univ Brest, Ifremer, CNRS, Biologie et Ecologie des Ecoystèmes marins profonds (BEEP)
J. Iñaki Guijarro: Université Paris Cité, CNRS UMR 3528
Ghislaine Henneke: Univ Brest, Ifremer, CNRS, Biologie et Ecologie des Ecoystèmes marins profonds (BEEP)
Ludovic Sauguet: Université Paris Cité, CNRS UMR 3528

Nature Communications, 2024, vol. 15, issue 1, 1-17

Abstract: Abstract Replication Protein A (RPA) plays a pivotal role in DNA replication by coating and protecting exposed single-stranded DNA, and acting as a molecular hub that recruits additional replication factors. We demonstrate that archaeal RPA hosts a winged-helix domain (WH) that interacts with two key actors of the replisome: the DNA primase (PriSL) and the replicative DNA polymerase (PolD). Using an integrative structural biology approach, combining nuclear magnetic resonance, X-ray crystallography and cryo-electron microscopy, we unveil how RPA interacts with PriSL and PolD through two distinct surfaces of the WH domain: an evolutionarily conserved interface and a novel binding site. Finally, RPA is shown to stimulate the activity of PriSL in a WH-dependent manner. This study provides a molecular understanding of the WH-mediated regulatory activity in central replication factors such as RPA, which regulate genome maintenance in Archaea and Eukaryotes.

Date: 2024
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DOI: 10.1038/s41467-024-55365-w

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