Binding to PCNA in Euryarchaeal DNA Replication requires two PIP motifs for DNA polymerase D and one PIP motif for DNA polymerase B

Benoît Castrec; Christophe Rouillon; Ghislaine Henneke; Didier Flament; Joël Querellou; Jean-Paul Raffin

doi:10.1016/j.jmb.2009.09.044

Binding to PCNA in Euryarchaeal DNA Replication requires two PIP motifs for DNA polymerase D and one PIP motif for DNA polymerase B

Benoît Castrec ¹ Christophe Rouillon ¹ Ghislaine Henneke ¹ Didier Flament ¹ Joël Querellou ¹ Jean-Paul Raffin ^{1, *}

* Auteur correspondant

1 LM2E - Laboratoire de microbiologie des environnements extrêmophiles

Abstract : Replicative DNA polymerases possess a canonical C-terminal proliferating cell nuclear antigen (PCNA)-binding motif termed the PCNA-interacting protein (PIP) box. We investigated the role of the PIP box on the functional interactions of the two DNA polymerases, PabPol B (family B) and PabPol D (family D), from the hyperthermophilic euryarchaeon Pyrococcus abyssi, with its cognate PCNA. The PIP box was essential for interactions of PabPol B with PCNA, as shown by surface plasmon resonance and primer extension studies. In contrast, binding of PabPol D to PCNA was affected only partially by removing the PIP motif. We identified a second palindromic PIP box motif at the N-terminus of the large subunit of PabPol D that was required for the interactions of PabPol D with PCNA. Thus, two PIP motifs were needed for PabPol D for binding to PabPCNA. Moreover, the C-terminus of PabPCNA was essential for stimulation of PabPol D activity but not for stimulation of PabPol B activity. Neither DNA polymerase interacted with the PabPCNA interdomain connecting loop. Our data suggest that distinct processes are involved in PabPol D and PabPol B binding to PCNA, raising the possibility that Archaea require two mechanisms for recruiting replicative DNA polymerases at the replication fork.

Keywords : DNA replication DNA polymerases PCNA-binding motifs PIP box Archaea

Type de document :

Article dans une revue

Journal of Molecular Biology, Elsevier, 2009, 394 (2), pp.209-218. 〈10.1016/j.jmb.2009.09.044〉

Domaine :

Sciences du Vivant [q-bio] / Microbiologie et Parasitologie

Liste complète des métadonnées

http://hal.univ-brest.fr/hal-00609928
Contributeur : Violaine Garguilo <>
Soumis le : mercredi 20 juillet 2011 - 15:12:39
Dernière modification le : jeudi 11 janvier 2018 - 06:16:39

Binding to PCNA in Euryarchaeal DNA Replication requires two PIP motifs for DNA polymerase D and one PIP motif for DNA polymerase B

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Binding to PCNA in Euryarchaeal DNA Replication requires two PIP motifs for DNA polymerase D and one PIP motif for DNA polymerase B

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