Strategy to discriminate between high and low affinity bindings of human immunodeficiency virus, type 1 integrase to viral DNA.
The last decade has contributed to our understanding of the three-dimensional structure of the human immunodeficiency virus, type 1 (HIV-1) integrase (IN) and to the description of how the enzyme catalyzes the viral DNA integration into the host DNA. Recognition of the viral DNA termini by IN is sequence-specific, and that of the host DNA does not require particular sequence, although in physicochemical studies IN fails to discriminate between the two interactions. Here, such discrimination was allowed thanks to a model system using designed oligonucleotides and peptides as binding structures. Spectroscopic (circular dichroism, NMR, and fluorescence anisotropy) techniques and biochemical (enzymatic and filter binding) assays clearly indicated that the amphipathic helix alpha4, located at the catalytic domain surface, is responsible for the specific high affinity binding of the enzyme to viral DNA. Analogues of the alpha4 peptide having increased helicity and still bearing the biologically relevant lysines 156 and 159 on the DNA binding face, and oligonucleotides conserving an intact attachment site, are required to achieve high affinity complexes (Kd of 1.5 nm). Data corroborate previous in vivo results obtained with mutated viruses.
Zargarian
Loussinée
L
Département de Biologie et Pharmacologie Structurales, UMR 8113 CNRS, Institut Gustave Roussy, Villejuif 94805 and Ecole Normale Supérieure de Cachan, France.
Benleumi
Mohamed Salah
MS
Renisio
Jean-Guillaume
JG
Merad
Hayate
H
Maroun
Richard G
RG
Wieber
Frédéric
F
Mauffret
Olivier
O
Porumb
Horea
H
Troalen
Frédéric
F
Fermandjian
Serge
S
D000595:Amino Acid Sequence; D001483:Base Sequence; D017931:DNA Primers; D004279:DNA, Viral / Q000378:metabolism*; D005454:Fluorescence Polarization; D019427:HIV Integrase / Q000378:metabolism*; D015497:HIV-1 / Q000201:enzymology*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D011487:Protein Conformation; D017386:Sequence Homology, Amino Acid; D012689:Sequence Homology, Nucleic Acid
D017931:DNA Primers; D004279:DNA, Viral; D019427:HIV Integrase