C-TERMINAL RETROVIRAL-TYPE ZINC FINGER DOMAIN FROM THE HIV-1 NUCLEOCAPSID PROTEIN IS STRUCTURALLY SIMILAR TO THE N-TERMINAL ZINC FINGER DOMAIN

Two-dimensional NMR spectroscopic and computational methods were employed for the structure determination of an 18-residue peptide with the amino acid sequence of the C-terminal retroviral-type (r.t.) zinc finger domain from the nucleocapsid protein (NCP) of HIV-1 [Zn(HIV1-F2)]. Unlike results obtained for the first retroviral-type zinc finger peptide, Zn(HIV1-F1), [Summers et al. (1990) Biochemistry 29, 329], broad signals indicative of conformational lability were observed in the 1H NMR spectrum of Zn(HIV1-F2) at 25-degrees-C. The NMR signals narrowed upon cooling to -2-degrees-C, enabling complete H-1 NMR signal assignment via standard two-dimensional (2D) NMR methods. Distance restraints obtained from qualitative analysis of 2D nuclear Overhauser effect (NOESY) data were used to generate 30 distance geometry (DG) structures with penalties (penalty = sum of the squared differences between interatomic distances defined in the restraints file and in the DG structures) in the range 0.02-0.03 angstrom2. All structures were qualitatively consistent with the experimental NOESY spectrum based on comparisons with 2D NOESY back-calculated spectra. Superposition of the backbone atoms (C, C-alpha, N) for residues C(1)-C(14) gave pairwise RMSD values in the range 0.16-0.75 angstrom. The folding of Zn(HIV1-F2) is very similar to that observed for Zn(HIV1-F1). Small differences observed between the two finger domains are localized to residues between His(9) and Cys(14), with residues M(11)-C(14) forming a 3(10) helical corner. Superposition of Zn(HIV1-F2) structures onto Zn(HIV1-FI) structures gave pairwise RMSD values in the ranges 0.4-0.7 angstrom [backbone atoms of residues C(1)-H(9) superpositioned] and 0.7-1.2 angstrom [backbone atoms of residues C(1)-C(14) superpositioned]. These results indicate that the r.t. zinc ringer sequences observed in retroviral NCPs, simple plant virus coat proteins, and in a human single-stranded nucleic acid binding protein share a common structural motif.