Aqueous solution structure of a hybrid lentiviral Tat peptide and a model of its interaction with HIV-1 TAR RNA.
Human immunodeficiency virus, type 1, (HIV-1) encodes a transactivating regulatory protein, called Tat, which is required for efficient transcription of the viral genome. Tat acts by binding to a specific RNA stem-loop element, called TAR, on nascent viral transcripts. The specificity of binding is principally determined by residues in a short, highly basic domain of Tat. The structure in aqueous solution of a biologically active peptide, comprised of the ten-amino acid HIV-1 Tat basic domain linked to a 15-amino acid segment of the core regulatory domain of another lentiviral Tat, i.e., that from equine infectious anemia virus (EIAV), has been determined. The restraint data set includes interproton distance bounds determined from two-dimensional nuclear Overhauser effect (2D NOE) spectra via a complete relaxation matrix analysis. Thirty structures consistent with the experimental data were generated via the distance geometry program DIANA. Subsequent restrained molecular mechanics calculations were used to define the conformational space subtended by the peptide. A large fraction of the 25-mer peptide assumes a structure in aqueous solution with the lysine- and arginine-rich HIV-1 basic domain being separated from the basic domain by a turn and characterized by a nascent helix as well. The Tat peptide/TAR complex could be modeled with the basic alpha-helix lying in the major groove of TAR such that important interactions of a putative specificity-endowing arginine are maintained and very slight widening of the major groove is entailed.
Mujeeb
A
A
Department of Pharmaceutical Chemistry, University of California, San Francisco 94143-0446, USA.
Parslow
T G
TG
Yuan
Y C
YC
James
T L
TL
D000595:Amino Acid Sequence; D000818:Animals; D015696:Gene Products, tat / Q000138:chemical synthesis / Q000737:chemistry* / Q000378:metabolism; D016325:HIV Long Terminal Repeat / Q000502:physiology*; D015497:HIV-1 / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D004860:Infectious Anemia Virus, Equine / Q000737:chemistry; D009682:Magnetic Resonance Spectroscopy; D008432:Mathematical Computing; D008958:Models, Molecular; D008969:Molecular Sequence Data; D017433:Protein Structure, Secondary; D017386:Sequence Homology, Amino Acid; D012996:Solutions; D054322:tat Gene Products, Human Immunodeficiency Virus
D015696:Gene Products, tat; D012996:Solutions; D054322:tat Gene Products, Human Immunodeficiency Virus