Binding to PCNA in Euryarchaeal DNA Replication Requires Two PIP Motifs for DNA Polymerase D and One PIP Motif for DNA Polymerase B

Type Article
Date 2009-11
Language English
Author(s) Castrec Benoit1, 2, Rouillon Christophe1, 2, Henneke Ghislaine1, 2, Flament Didier1, 2, Querellou Joel1, 2, Raffin Jean-Paul2, 3
Affiliation(s) 1 : Univ Bretagne Occidentale, UMR 6197, Lab Microbiol Environm Extremes, F-29280 Plouzane, France.
2 : IFREMER, UMR 6197, Lab Microbiol Environm Extremes, F-29280 Plouzane, France.
3 : CNRS, UMR 6197, Lab Microbiol Environm Extremes, F-29280 Plouzane, France.
Source Journal of Molecular Biology (0022-2836) (Elsevier), 2009-11 , Vol. 394 , N. 2 , P. 209-218
DOI 10.1016/j.jmb.2009.09.044
WOS© Times Cited 16
Keyword(s) Archaea, PIP box, PCNA binding motifs, DNA polymerases, DNA replication
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. (C) 2009 Elsevier Ltd. All rights reserved.
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Castrec Benoit, Rouillon Christophe, Henneke Ghislaine, Flament Didier, Querellou Joel, Raffin Jean-Paul (2009). Binding to PCNA in Euryarchaeal DNA Replication Requires Two PIP Motifs for DNA Polymerase D and One PIP Motif for DNA Polymerase B. Journal of Molecular Biology, 394(2), 209-218. Publisher's official version : https://doi.org/10.1016/j.jmb.2009.09.044 , Open Access version : https://archimer.ifremer.fr/doc/00000/7317/