Modulation of the Pyrococcus abyssi NucS endonuclease activity by replication clamp at functional and structural levels.

Christophe Creze 1 Alessio Ligabue 2 Sébastien Laurent 1 Roxane Lestini 2 Sergey Laptenok 2 Joelle Khun 2 Marten Vos 2 Mirjam Czjzek 3 Hannu Myllykallio 2 Didier Flament 1
1 LM2E - Laboratoire de microbiologie des environnements extrêmophiles
2 LOB - Laboratoire d'optique et biosciences
3 UMR7139 - Végétaux marins et biomolécules
Abstract : Pyrococcus abyssi NucS is the founding member of a new family of structure-specific DNA endonucleases that interact with the replication clamp proliferating cell nuclear antigen (PCNA). Using a combination of small angle x-ray scattering and surface plasmon resonance analyses, we demonstrate the formation of a stable complex in solution, in which one molecule of the PabNucS homodimer binds to the outside surface of the PabPCNA homotrimer. Using fluorescent labels, PCNA is shown to increase the binding affinity of NucS toward single-strand/double-strand junctions on 5' and 3' flaps, as well as to modulate the cleavage specificity on the branched DNA structures. Our results indicate that the presence of a single major contact between the PabNucS and PabPCNA proteins, together with the complex-induced DNA bending, facilitate conformational flexibility required for specific cleavage at the single-strand/double-strand DNA junction.
Keywords : CELL NUCLEAR ANTIGEN SMALL-ANGLE SCATTERING DNA-LIGASE-I RESONANCE ENERGY-TRANSFER RAY SOLUTION SCATTERING DOUBLE-STRANDED DNA RECBCD ENZYME X-RAY HYPERTHERMOPHILIC ARCHAEON CRYSTAL-STRUCTURE
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Article dans une revue
Journal of Biological Chemistry, American Society for Biochemistry and Molecular Biology, 2012, 287 (19), pp.15648-60. 〈10.1074/jbc.M112.346361〉
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Soumis le : vendredi 5 avril 2013 - 15:39:45
Dernière modification le : jeudi 10 mai 2018 - 02:08:16

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Christophe Creze, Alessio Ligabue, Sébastien Laurent, Roxane Lestini, Sergey Laptenok, et al.. Modulation of the Pyrococcus abyssi NucS endonuclease activity by replication clamp at functional and structural levels.. Journal of Biological Chemistry, American Society for Biochemistry and Molecular Biology, 2012, 287 (19), pp.15648-60. 〈10.1074/jbc.M112.346361〉. 〈hal-00808519〉

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