title: Role of the PWWP domain of lens epithelium-derived growth factor (LEDGF)/p75 cofactor in lentiviral integration targeting.
abstract: LEDGF/p75 is a chromatin-interacting, cellular cofactor of HIV integrase that dictates lentiviral integration site preference. In this study we determined the role of the PWWP domain of LEDGF/p75 in tethering and targeting of the lentiviral pre-integration complex, employing potent knockdown cell lines allowing analysis in the absence of endogenous LEDGF/p75. Deletion of the PWWP domain resulted in a diffuse subnuclear distribution pattern, loss of interaction with condensed chromatin, and failure to rescue proviral integration, integration site distribution, and productive virus replication. Substitution of the PWWP domain of LEDGF/p75 with that of hepatoma-derived growth factor or HDGF-related protein-2 rescued viral replication and lentiviral integration site distribution in LEDGF/p75-depleted cells. Replacing all chromatin binding elements of LEDGF/p75 with full-length hepatoma-derived growth factor resulted in more integration in genes combined with a preference for CpG islands. In addition, we showed that any PWWP domain targets SMYD1-like sequences. Analysis of integration preferences of lentiviral vectors for epigenetic marks indicates that the PWWP domain is critical for interactions specifying the relationship of integration sites to regions enriched in specific histone post-translational modifications.
authors: Division of Molecular Medicine, KU Leuven, 3000 Leuven, Belgium. Electronic address: rik.gijsbers@med.kuleuven.be.
authors: Rik
authors: R
authors: Gijsbers
authors: Division of Molecular Medicine, KU Leuven, 3000 Leuven, Belgium.
authors: Sofie
authors: S
authors: Vets
authors: Division of Molecular Medicine, KU Leuven, 3000 Leuven, Belgium.
authors: Jan
authors: J
authors: De Rijck
authors: Department of Microbiology, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania 19104.
authors: Karen E
authors: KE
authors: Ocwieja
authors: Department of Microbiology, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania 19104.
authors: Keshet
authors: K
authors: Ronen
authors: Department of Microbiology, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania 19104.
authors: Nirav
authors: N
authors: Malani
authors: Department of Microbiology, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania 19104.
authors: Frederic D
authors: FD
authors: Bushman
authors: Division of Molecular Medicine, KU Leuven, 3000 Leuven, Belgium. Electronic address: zeger.debyser@med.kuleuven.be.
authors: Zeger
authors: Z
authors: Debyser
mesh_terms: D048868:Adaptor Proteins, Signal Transducing / Q000235:genetics / Q000378:metabolism*; D000595:Amino Acid Sequence; D002843:Chromatin; D018899:CpG Islands / Q000502:physiology; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D015497:HIV-1 / Q000235:genetics / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D009124:Muscle Proteins / Q000235:genetics / Q000378:metabolism; D017434:Protein Structure, Tertiary; D017384:Sequence Deletion; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism*; D016662:Virus Integration / Q000502:physiology*
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D002843:Chromatin; D004268:DNA-Binding Proteins; D009124:Muscle Proteins; C504649:PSIP1 protein, human; C484340:SMYD1 protein, human; D014157:Transcription Factors