Three-dimensional 1H NMR structure of the nucleocapsid protein NCp10 of Moloney murine leukemia virus.
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 > or = 5, NCp10 binds one zinc atom and the complexation induces a folding of the CysX2CysX4HEsX4Cys 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 A for backbone atoms and 1.709 +/- 0.187 A 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.
Départemente de Pharmacochimie Moléculaire et Structurale, U266 INSERM-URA D 1500 CNRS, Faculté de Pharmacie, Paris, France.
H
H
Déméné
N
N
Jullian
N
N
Morellet
H
H
de Rocquigny
F
F
Cornille
B
B
Maigret
B P
BP
Roques
D000595:Amino Acid Sequence; D017124:Conserved Sequence; D015683:Gene Products, gag / Q000138:chemical synthesis / Q000737:chemistry*; D006859:Hydrogen; D009682:Magnetic Resonance Spectroscopy / Q000379:methods; D008433:Mathematics; D008958:Models, Molecular; D008969:Molecular Sequence Data; D008979:Moloney murine leukemia virus / Q000737:chemistry*; D011487:Protein Conformation; D014758:Viral Core Proteins / Q000138:chemical synthesis / Q000737:chemistry*; D016335:Zinc Fingers
D015683:Gene Products, gag; D014758:Viral Core Proteins; C068662:retroviral nucleocapsid protein NCp10, Moloney murine leukemia virus; D006859:Hydrogen