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

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.
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https://hal.univ-brest.fr/hal-00609913
Contributor : Violaine Garguilo <>
Submitted on : Wednesday, July 20, 2011 - 2:54:25 PM
Last modification on : Thursday, January 11, 2018 - 6:16:39 AM

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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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