The glycine-rich motif of Pyrococcus abyssi DNA polymerase D is critical for protein stability.

Benoît Castrec 1 Sébastien Laurent 1 Ghislaine Henneke 1, * Didier Flament 1 Jean-Paul Raffin 1
* Auteur correspondant
1 LM2E - Laboratoire de microbiologie des environnements extrêmophiles
Abstract : A glycine-rich motif described as being involved in human polymerase delta proliferating cell nuclear antigen (PCNA) binding has also been identified in all euryarchaeal DNA polymerase D (Pol D) family members. We redefined the motif as the (G)-PYF box. In the present study, Pol D (G)-PYF box motif mutants from Pyrococcus abyssi were generated to investigate its role in functional interactions with the cognate PCNA. We demonstrated that this motif is not essential for interactions between PabPol D (P. abyssi Pol D) and PCNA, using surface plasmon resonance and primer extension studies. Interestingly, the (G)-PYF box is located in a hydrophobic region close to the active site. The (G)-PYF box mutants exhibited altered DNA binding properties. In addition, the thermal stability of all mutants was reduced compared to that of wild type, and this effect could be attributed to increased exposure of the hydrophobic region. These studies suggest that the (G)-PYF box motif mediates intersubunit interactions and that it may be crucial for the thermostability of PabPol D.
Keywords : DNA replication DNA polymerase D (G)-PYF box Archaea thermostability
Type de document :
Article dans une revue
Journal of Molecular Biology, Elsevier, 2010, 396 (4), pp.840-848. 〈10.1016/j.jmb.2010.01.006〉
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http://hal.univ-brest.fr/hal-00609913
Contributeur : Violaine Garguilo <>
Soumis le : mercredi 20 juillet 2011 - 14:54:25
Dernière modification le : jeudi 11 janvier 2018 - 06:16:39

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Benoît Castrec, Sébastien Laurent, Ghislaine Henneke, Didier Flament, Jean-Paul Raffin. The glycine-rich motif of Pyrococcus abyssi DNA polymerase D is critical for protein stability.. Journal of Molecular Biology, Elsevier, 2010, 396 (4), pp.840-848. 〈10.1016/j.jmb.2010.01.006〉. 〈hal-00609913〉

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