The Glycine-Rich Motif of Pyrococcus abyssi DNA Polymerase D Is Critical for Protein Stability
| Autor: | Benoît Castrec, Jean-Paul Raffin, Ghislaine Henneke, Sébastien Laurent, Didier Flament |
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| Přispěvatelé: | Laboratoire de microbiologie des environnements extrêmophiles (LM2E), Centre National de la Recherche Scientifique (CNRS)-Université de Brest (UBO)-Institut Français de Recherche pour l'Exploitation de la Mer (IFREMER) |
| Rok vydání: | 2010 |
| Předmět: |
Pyrococcus abyssi
MESH: Enzyme Stability MESH: Sequence Homology Amino Acid DNA polymerase MESH: DNA Polymerase III Amino Acid Motifs Mutant MESH: Catalytic Domain MESH: Amino Acid Sequence MESH: Base Sequence Primer extension MESH: Recombinant Proteins MESH: Amino Acid Motifs chemistry.chemical_compound Structural Biology Catalytic Domain Enzyme Stability MESH: Pyrococcus abyssi Conserved Sequence Polymerase 0303 health sciences MESH: Conserved Sequence biology MESH: DNA Archaeal MESH: Hydrophobic and Hydrophilic Interactions 030302 biochemistry & molecular biology MESH: Archaeal Proteins (G)-PYF box MESH: Fluorescent Dyes MESH: Amino Acid Substitution Recombinant Proteins MESH: Glycine MESH: Surface Plasmon Resonance MESH: Mutagenesis Site-Directed DNA Archaeal [SDV.MP]Life Sciences [q-bio]/Microbiology and Parasitology Biochemistry Hydrophobic and Hydrophilic Interactions Archaeal Proteins Molecular Sequence Data Glycine DNA replication 03 medical and health sciences Proliferating Cell Nuclear Antigen Oxazines Protein Interaction Domains and Motifs Amino Acid Sequence DNA polymerase D (G)-PYF box Molecular Biology DNA Polymerase III Fluorescent Dyes 030304 developmental biology MESH: Protein Interaction Domains and Motifs MESH: Molecular Sequence Data Base Sequence Sequence Homology Amino Acid Surface Plasmon Resonance biology.organism_classification Archaea Molecular biology thermostability Proliferating cell nuclear antigen MESH: Proliferating Cell Nuclear Antigen Amino Acid Substitution chemistry Mutagenesis Site-Directed biology.protein DNA polymerase D MESH: Oxazines DNA |
| Zdroj: | Journal of Molecular Biology Journal of Molecular Biology, Elsevier, 2010, 396 (4), pp.840-848. ⟨10.1016/j.jmb.2010.01.006⟩ Journal Of Molecular Biology (0022-2836) (Academic Press Ltd-Elsevier Science Ltd), 2010-03, Vol. 396, N. 4, P. 840-848 |
| ISSN: | 0022-2836 1089-8638 |
| DOI: | 10.1016/j.jmb.2010.01.006 |
| Popis: | En libre-accès sur Archimer : http://archimer.ifremer.fr/doc/00002/11293/7879.pdf; International audience; 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. |
| Databáze: | OpenAIRE |
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