NMR solution structure of the RNA-binding peptide from human immunodeficiency virus (type 1) Rev.
NMR spectroscopy has been used to solve the three-dimensional solution structure of a minimal RNA-binding domain of the Rev protein from the human immunodeficiency virus (type 1), an essential regulatory protein for viral replication. The presence of 10 arginine residues in the 17-residue peptide Rev34-50 caused significant problems in assignment of the NMR spectra. To improve spectral resolution, the peptide was synthesized with an alanine replacing a nonessential arginine and with selectively 15N-labeled residues. Contrary to Chou-Fasman modeling predictions an alpha-helix was detected in both water and 20% trifluoroethanol (TFE) and was found to span residues that constitute the RNA-binding and nuclear-localizing domains of Rev. The sequence-specific information provided by the NMR data gives a full description of the solution conformation of Rev34-50 which serves as a template for investigating binding of the peptide to RNA from the Rev response element (RRE). Preliminary modeling suggests that the helix can fit neatly into the expanded major groove of the RRE where interactions between the peptide side chains and the RNA can be identified. These data may aid the construction of a suitable pharmacophore model for the rational design of molecules that block Rev-RNA binding and inhibit HIV replication.
Scanlon
M J
MJ
Centre for Drug Design and Development, University of Queensland, St Lucia, Australia.
Fairlie
D P
DP
Craik
D J
DJ
Englebretsen
D R
DR
West
M L
ML
D000595:Amino Acid Sequence; D018360:Crystallography, X-Ray; D015694:Gene Products, rev / Q000737:chemistry*; D015497:HIV-1 / Q000378:metabolism*; D009682:Magnetic Resonance Spectroscopy / Q000379:methods; D008961:Models, Structural; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry; D010455:Peptides / Q000138:chemical synthesis / Q000737:chemistry; D011487:Protein Conformation; D016601:RNA-Binding Proteins / Q000737:chemistry*; D054321:rev Gene Products, Human Immunodeficiency Virus
D015694:Gene Products, rev; D010446:Peptide Fragments; D010455:Peptides; D016601:RNA-Binding Proteins; D054321:rev Gene Products, Human Immunodeficiency Virus