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Three-dimensional 1H NMR structure of the nucleocapsid protein NCp10 of Moloney murine leukemia virus

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Summary

The nucleocapsid protein of Moloney murine leukemia virus (NCp10) is a 56-amino acid protein which contains one zinc finger of the CysX2CysX4HisX4Cys form, a highly conserved motif present in most retroviruses and retroelements. At pH≥5, NCp10 binds one zinc atom and the complexation induces a folding of the CysX2CysX4HisX4Cys box, similar to that observed for the zinc-binding domains of HIV-1 NC protein. The three-dimensional structure of NCp10 has been determined in aqueous solution by 600 MHz 1H NMR spectroscopy. The proton resonances could be almost completely assigned by means of phase-sensitive double-quantum-filtered COSY, TOCSY and NOESY techniques. NOESY spectra yielded 597 relevant structural constraints, which were used as input for distance geometry calculations with DIANA. Further refinement was performed by minimization with the program AMBER, which was modified by introducing a zinc force field. The solution structure is characterized by a well-defined central zinc finger (rmsd of 0.747±0.209 Å for backbone atoms and 1.709±0.187 Å when all atoms are considered), surrounded by flexible N- and C-terminal domains. The Tyr28, Trp35, Lys37, Lys41 and Lys42 residues, which are essential for activity, lie on the same face of the zinc finger, forming a bulge structure probably involved in viral RNA binding. The significance of these structural characteristics for the various biological functions of the protein is discussed, taking into account the results obtained with various mutants.

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Authors and Affiliations

  1. Département de Pharmacochimie Moléculaire et Structurale, U266 INSERM-URA D 1500 CNRS, Faculté de Pharmacie, 4, Avenue de l'Observatoire, 75270, Paris Cedex 06, France

    Hélène Déméné, Nathalie Jullian, Nelly Morellet, Hugues de Rocquigny, Fabrice Cornille & Bernard P. Roques

  2. Laboratoire Chimie Théorique, Université Nancy I, B.P. 239, 54506, Vandoeuvre-lés-Nancy Cedex, France

    Bernard Maigret

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  1. Hélène Déméné
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  2. Nathalie Jullian
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  3. Nelly Morellet
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  4. Hugues de Rocquigny
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  5. Fabrice Cornille
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  6. Bernard Maigret
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  7. Bernard P. Roques
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Déméné, H., Jullian, N., Morellet, N. et al. Three-dimensional 1H NMR structure of the nucleocapsid protein NCp10 of Moloney murine leukemia virus. J Biomol NMR 4, 153–170 (1994). https://doi.org/10.1007/BF00175244

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  • DOI: https://doi.org/10.1007/BF00175244

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