title: Altering murine leukemia virus integration through disruption of the integrase and BET protein family interaction.
abstract: We report alterations to the murine leukemia virus (MLV) integrase (IN) protein that successfully result in decreasing its integration frequency at transcription start sites and CpG islands, thereby reducing the potential for insertional activation. The host bromo and extraterminal (BET) proteins Brd2, 3 and 4 interact with the MLV IN protein primarily through the BET protein ET domain. Using solution NMR, protein interaction studies, and next generation sequencing, we show that the C-terminal tail peptide region of MLV IN is important for the interaction with BET proteins and that disruption of this interaction through truncation mutations affects the global targeting profile of MLV vectors. The use of the unstructured tails of gammaretroviral INs to direct association with complexes at active promoters parallels that used by histones and RNA polymerase II. Viruses bearing MLV IN C-terminal truncations can provide new avenues to improve the safety profile of gammaretroviral vectors for human gene therapy.
authors.affiliation: Department of Pharmacology, Robert Wood Johnson Medical School, Rutgers University, 675 Hoes Lane, Piscataway, NJ 08854, USA.
authors.forename: Sriram
authors.initials: S
authors.lastname: Aiyer
authors.affiliation: Center for Advanced Biotechnology and Medicine, Department of Molecular Biology and Biochemistry, and Northeast Structural Genomics Consortium, Rutgers, The State University of New Jersey, 679 Hoes Lane West Piscataway, NJ 08854, USA.
authors.forename: G V T
authors.initials: GV
authors.lastname: Swapna
authors.affiliation: Department of Microbiology, Perelman School of Medicine, University of Pennsylvania, 3610 Hamilton Walk, Philadelphia, PA 19104, USA.
authors.forename: Nirav
authors.initials: N
authors.lastname: Malani
authors.affiliation: Center for Advanced Biotechnology and Medicine, Department of Molecular Biology and Biochemistry, and Northeast Structural Genomics Consortium, Rutgers, The State University of New Jersey, 679 Hoes Lane West Piscataway, NJ 08854, USA.
authors.forename: James M
authors.initials: JM
authors.lastname: Aramini
authors.affiliation: Department of Biochemistry, Robert Wood Johnson Medical School, UMDNJ, 675 Hoes Lane, Piscataway, NJ 08854, USA.
authors.forename: William M
authors.initials: WM
authors.lastname: Schneider
authors.affiliation: Center for Retrovirus Research and College of Pharmacy, The Ohio State University, 484 W. 12th Ave., 508 Riffe Building, Columbus, OH 43210, USA.
authors.forename: Matthew R
authors.initials: MR
authors.lastname: Plumb
authors.affiliation: Department of Biology, Brooklyn College, 417 Ingersoll Extension and the Graduate Center of the City University of New York, Brooklyn, NY 11210, USA.
authors.forename: Mustafa
authors.initials: M
authors.lastname: Ghanem
authors.affiliation: Center for Retrovirus Research and College of Pharmacy, The Ohio State University, 484 W. 12th Ave., 508 Riffe Building, Columbus, OH 43210, USA.
authors.forename: Ross C
authors.initials: RC
authors.lastname: Larue
authors.affiliation: Center for Retrovirus Research and College of Pharmacy, The Ohio State University, 484 W. 12th Ave., 508 Riffe Building, Columbus, OH 43210, USA.
authors.forename: Amit
authors.initials: A
authors.lastname: Sharma
authors.affiliation: Department of Biology, Brooklyn College, 417 Ingersoll Extension and the Graduate Center of the City University of New York, Brooklyn, NY 11210, USA.
authors.forename: Barbara
authors.initials: B
authors.lastname: Studamire
authors.affiliation: Center for Retrovirus Research and College of Pharmacy, The Ohio State University, 484 W. 12th Ave., 508 Riffe Building, Columbus, OH 43210, USA.
authors.forename: Mamuka
authors.initials: M
authors.lastname: Kvaratskhelia
authors.affiliation: Department of Microbiology, Perelman School of Medicine, University of Pennsylvania, 3610 Hamilton Walk, Philadelphia, PA 19104, USA.
authors.forename: Frederic D
authors.initials: FD
authors.lastname: Bushman
authors.affiliation: Center for Advanced Biotechnology and Medicine, Department of Molecular Biology and Biochemistry, and Northeast Structural Genomics Consortium, Rutgers, The State University of New Jersey, 679 Hoes Lane West Piscataway, NJ 08854, USA Department of Biochemistry and Molecular Biology, Robert Wood Johnson Medical School, Rutgers, The State University of New Jersey, 675 Hoes Lane, Piscataway, NJ 08854, USA roth@rwjms.rutgers.edu.
authors.forename: Gaetano T
authors.initials: GT
authors.lastname: Montelione
authors.affiliation: Department of Pharmacology, Robert Wood Johnson Medical School, Rutgers University, 675 Hoes Lane, Piscataway, NJ 08854, USA Department of Biochemistry and Molecular Biology, Robert Wood Johnson Medical School, Rutgers, The State University of New Jersey, 675 Hoes Lane, Piscataway, NJ 08854, USA roth@rwjms.rutgers.edu.
authors.forename: Monica J
authors.initials: MJ
authors.lastname: Roth
mesh_terms: D000595:Amino Acid Sequence; D001665:Binding Sites; D018899:CpG Islands; D057809:HEK293 Cells; D059014:High-Throughput Nucleotide Sequencing; D006801:Humans; D019426:Integrases / Q000737:chemistry*; D009052:Leukemia Virus, Murine / Q000235:genetics*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D019906:Nuclear Magnetic Resonance, Biomolecular; D011485:Protein Binding; D054730:Protein Interaction Domains and Motifs; D016601:RNA-Binding Proteins / Q000737:chemistry*; D017422:Sequence Analysis, DNA; D017384:Sequence Deletion; D014157:Transcription Factors; D024363:Transcription Initiation Site; D014764:Viral Proteins / Q000737:chemistry*; D016662:Virus Integration; D000092468:Bromodomain Containing Proteins
chemical_list: D019426:Integrases; D016601:RNA-Binding Proteins; D014157:Transcription Factors; D014764:Viral Proteins; C109106:BRD3 protein, human; D000092468:Bromodomain Containing Proteins