title: Peptides derived from HIV-1 Rev inhibit HIV-1 integrase in a shiftide mechanism.
abstract: The HIV-1 Integrase protein (IN) mediates the integration of the viral cDNA into the host genome. IN is an emerging target for anti-HIV drug design, and the first IN-inhibitor was recently approved by the FDA. We have developed a new approach for inhibiting IN by "shiftides": peptides derived from its cellular binding protein LEDGF/p75 that inhibit IN by shifting its oligomerization equilibrium from the active dimer to an inactive tetramer. In addition, we described two peptides derived from the HIV-1 Rev protein that interact with IN and inhibit its activity in vitro and in cells. In the current study, we show that the Rev-derived peptides also act as shiftides. Analytical gel filtration and cross-linking experiments showed that IN was dimeric when bound to the viral DNA, but tetrameric in the presence of the Rev-derived peptides. Fluorescence anisotropy studies revealed that the Rev-derived peptides inhibited the DNA binding of IN. The Rev-derived peptides inhibited IN catalytic activity in vitro in a concentration-dependent manner. Inhibition was much more significant when the peptides were added to free IN before it bound the viral DNA than when the peptides were added to a preformed IN-DNA complex. This confirms that the inhibition is due to the ability of the peptides to shift the oligomerization equilibrium of the free IN toward a tetramer that binds much weaker to the viral DNA. We conclude that protein-protein interactions of IN may serve as a general valuable source for shiftide design.
authors.affiliation: Institute of Chemistry, Department of Biological Chemistry, The Hebrew University of Jerusalem, Givat Ram, Jerusalem 91904, Israel. zvi.hayouka@mail.huji.ac.il
authors.forename: Zvi
authors.initials: Z
authors.lastname: Hayouka
authors.forename: Joseph
authors.initials: J
authors.lastname: Rosenbluh
authors.forename: Aviad
authors.initials: A
authors.lastname: Levin
authors.forename: Michal
authors.initials: M
authors.lastname: Maes
authors.forename: Abraham
authors.initials: A
authors.lastname: Loyter
authors.forename: Assaf
authors.initials: A
authors.lastname: Friedler
mesh_terms: D000595:Amino Acid Sequence; D002384:Catalysis / Q000187:drug effects; D004279:DNA, Viral / Q000494:pharmacology; D019427:HIV Integrase / Q000378:metabolism*; D008954:Models, Biological; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry* / Q000494:pharmacology*; D011485:Protein Binding / Q000187:drug effects; D011487:Protein Conformation / Q000187:drug effects*; D020836:Protein Structure, Quaternary; D020079:Terminal Repeat Sequences / Q000235:genetics; D054321:rev Gene Products, Human Immunodeficiency Virus / Q000737:chemistry*
chemical_list: D004279:DNA, Viral; D010455:Peptides; D054321:rev Gene Products, Human Immunodeficiency Virus; C514860:rev protein, Human Immunodeficiency Virus-1; D019427:HIV Integrase; C514801:p31 integrase protein, Human immunodeficiency virus 1