LEDGF/p75 interacts with divergent lentiviral integrases and modulates their enzymatic activity in vitro.
Transcriptional co-activator LEDGF/p75 is the major cellular interactor of HIV-1 integrase (IN), critical to efficient viral replication. In this work, a series of INs from the Betaretrovirus, Gammaretrovirus, Deltaretrovirus, Spumavirus and Lentivirus retroviral genera were tested for interaction with the host factor. None of the non-lentiviral INs possessed detectable affinity for LEDGF in either pull-down or yeast two-hybrid assays. In contrast, all lentiviral INs examined, including those from bovine immunodeficiency virus (BIV), maedi-visna virus (MVV) and equine infectious anemia virus (EIAV) readily interacted with LEDGF. Mutation of Asp-366 to Asn in LEDGF ablated the interaction, suggesting a common mechanism of the host factor recognition by the INs. LEDGF potently stimulated strand transfer activity of divergent lentiviral INs in vitro. Unprecedentedly, in the presence of the host factor, EIAV IN almost exclusively catalyzed concerted integration, whereas HIV-1 IN promoted predominantly half-site integration, and BIV IN was equally active in both types of strand transfer. Concerted BIV and EIAV integration resulted in 5 bp duplications of the target DNA sequences. These results confirm that the interaction with LEDGF is conserved within and limited to Lentivirus and strongly argue that the host factor is intimately involved in the catalysis of lentiviral DNA integration.
Cherepanov
Peter
P
Section of Infectious Diseases, Division of Medicine, Imperial College London, St Mary's Campus, Norfolk Place, W2 1PG, London, UK. p.cherepanov@imperial.ac.uk
D048868:Adaptor Proteins, Signal Transducing / Q000737:chemistry / Q000378:metabolism*; D000818:Animals; D002417:Cattle; D004279:DNA, Viral / Q000378:metabolism; D004789:Enzyme Activation; D006736:Horses; D006801:Humans; D016087:Immunodeficiency Virus, Bovine / Q000201:enzymology; D004860:Infectious Anemia Virus, Equine / Q000201:enzymology; D019426:Integrases / Q000378:metabolism*; D036341:Intercellular Signaling Peptides and Proteins / Q000737:chemistry / Q000378:metabolism; D016086:Lentivirus / Q000201:enzymology*; D012190:Retroviridae / Q000201:enzymology; D017422:Sequence Analysis, DNA; D014157:Transcription Factors / Q000737:chemistry / Q000378:metabolism*; D014764:Viral Proteins / Q000378:metabolism*
D048868:Adaptor Proteins, Signal Transducing; D004279:DNA, Viral; D036341:Intercellular Signaling Peptides and Proteins; C504649:PSIP1 protein, human; D014157:Transcription Factors; D014764:Viral Proteins; C119380:lens epithelium-derived growth factor; D019426:Integrases