HIV-1 integrase crosslinked oligomers are active in vitro

Faure, A.

doi:10.1093/nar/gki241

Cited by 174 publications

(222 citation statements)

References 42 publications

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“…Inhibition of IN is achieved before its binding to the full-length LEDGF/p75 and the tethering to the chromosomes, which takes place after nuclear import. Moreover, because the IN tetramer also is unable to bind directly to the processed DNA as shown by cross-linking experiments (12), shifting the oligomeric state of IN toward a tetramer inhibits the strand transfer of a processed DNA template. In summary, the shiftide approach results in inhibition of both integration steps, making it advantageous over strand-transfer inhibitors, which inhibit only the second integration step.…”

Section: Discussionmentioning

confidence: 99%

“…The integration proceeds by two steps (11): (i) 3Ј end processing, in which IN creates the DNA template for integration by removing dinucleotides from the 3Ј ends of both ends of the viral DNA LTRs after reverse transcription in the cytoplasm; and (ii) strand transfer, which is, after nuclear import, integration of the viral DNA template into the target host DNA. IN is in equilibrium among dimeric, tetrameric, and high-order oligomeric states (12)(13)(14). Dimeric IN binds at each end of the viral DNA during the 3Ј end processing in the cytoplasm (15).…”

mentioning

confidence: 99%

“…After nuclear import, the two LTR DNA-bound dimers approach each other in the presence of the cellular protein lens epithelium-derived growth factor (LEDGF)/p75 and form a tetramer, and the integration proceeds to the strand-transfer step (16). The free IN tetramer does not bind DNA directly, and tetramerization occurs only by the interaction between two DNA-bound IN dimers (12). Incorrect oligomerization of IN in terms of time and localization may prevent the essential native assembly of its complexes with the viral DNA LTR ends (17).…”

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confidence: 99%

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Inhibiting HIV-1 integrase by shifting its oligomerization equilibrium

Hayouka

Rosenbluh²,

Levin³

et al. 2007

Proc. Natl. Acad. Sci. U.S.A.

172

197

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Proteins are involved in various equilibria that play a major role in their activity or regulation. The design of molecules that shift such equilibria is of great therapeutic potential. This fact was demonstrated in the cases of allosteric inhibitors, which shift the equilibrium between active and inactive (R and T) states, and chemical chaperones, which shift folding equilibrium of proteins. Here, we expand these concepts and propose the shifting of oligomerization equilibrium of proteins as a general methodology for drug design. We present a strategy for inhibiting proteins by ''shiftides'': ligands that specifically bind to an inactive oligomeric state of a disease-related protein and modulate its activity by shifting the oligomerization equilibrium of the protein toward it. We demonstrate the feasibility of our approach for the inhibition of the HIV-1 integrase (IN) protein by using peptides derived from its cellularbinding protein, LEDGF/p75, which specifically inhibit IN activity by a noncompetitive mechanism. The peptides inhibit the DNA-binding of IN by shifting the IN oligomerization equilibrium from the active dimer toward the inactive tetramer, which is unable to catalyze the first integration step of 3 end processing. The LEDGF/ p75-derived peptides inhibit the enzymatic activity of IN in vitro and consequently block HIV-1 replication in cells because of the lack of integration. These peptides are promising anti-HIV lead compounds that modulate oligomerization of IN via a previously uncharacterized mechanism, which bears advantages over the conventional interface dimerization inhibitors.allostery ͉ protein equilibrium ͉ shiftides ͉ peptides ͉ drug design

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Section: Discussionmentioning

confidence: 99%

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confidence: 99%

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confidence: 99%

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Inhibiting HIV-1 integrase by shifting its oligomerization equilibrium

Hayouka

Rosenbluh²,

Levin³

et al. 2007

Proc. Natl. Acad. Sci. U.S.A.

172

197

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“…Alternatively, in the second study, p75 appeared to interfere with higher order oligomerization of recombinant HIV IN [104]. Since achieving the full-site reaction in vitro is thought to require the tetrameric form of the enzyme while the dimer mediates half-site integration [154,157,158], properly coupled joining may require that p75 binding occur after assembly of a mature "synaptic complex" in which tetrameric HIV-1 IN is complexed to the viral cDNA ends [104]. A note of complexity was added by Pandey et al, who found that full site integration by HIV IN was promoted at lower p75 concentrations but inhibited at higher concentrations [103].…”

Section: Modeling P75 Function In the Hiv Life Cyclementioning

confidence: 98%

“…Some IN residues located in the pocket at the p75-IN dimer interface are highlighted at the bottom of the three domain illustration, with those N-terminal to E152 in one monomer, and those C-terminal to E152 in the other monomer of the IN dimer. Although its exact oligomeric state and abundance in the PIC is still ambiguous, a wealth of in vitro enzymatic and virological complementation studies suggest IN functions to integrate both long terminal repeat (LTR) ends into host DNA in a coupled (concerted) fashion, and that it acts as a multimer, with a tetramer likely [73,154,157,158,[188][189][190][191][192]. In vitro, an IN dimer is sufficient for 3′ end processing but a tetramer appears needed for DNA strand transfer activity [154,157,158].…”

Section: Targeting Lentiviral Vectorsmentioning

confidence: 99%

Integrase, LEDGF/p75 and HIV replication

Poeschla

2008

Cell. Mol. Life Sci.

113

101

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HIV integrates a DNA copy of its genome into a host cell chromosome in each replication cycle. The essential DNA cleaving and joining chemistry of integration is known, but there is less understanding of the process as it occurs in a cell, where two complex and dynamic macromolecular entities are joined: the viral pre-integration complex and chromatin. Among implicated cellular factors, much recent attention has coalesced around LEDGF/p75, a nuclear protein that may act as a chromatin docking factor or receptor for lentiviral pre-integration complexes. LEDGF/p75 tethers HIV integrase to chromatin, protects it from degradation, and strongly influences the genome-wide pattern of HIV integration. Depleting the protein from cells and/or over-expressing its integrase-binding domain blocks viral replication. Current goals are to establish the underlying mechanisms and to determine whether this knowledge can be exploited for antiviral therapy or for targeting lentiviral vector integration in human gene therapy.

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