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Nucleic Acids Res. 2014 May;42(9):5917-28. doi: 10.1093/nar/gku175. Epub 2014 Mar 12.

Altering murine leukemia virus integration through disruption of the integrase and BET protein family interaction.

Author information

1
Department of Pharmacology, Robert Wood Johnson Medical School, Rutgers University, 675 Hoes Lane, Piscataway, NJ 08854, USA.
2
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.
3
Department of Microbiology, Perelman School of Medicine, University of Pennsylvania, 3610 Hamilton Walk, Philadelphia, PA 19104, USA.
4
Department of Biochemistry, Robert Wood Johnson Medical School, UMDNJ, 675 Hoes Lane, Piscataway, NJ 08854, USA.
5
Center for Retrovirus Research and College of Pharmacy, The Ohio State University, 484 W. 12th Ave., 508 Riffe Building, Columbus, OH 43210, USA.
6
Department of Biology, Brooklyn College, 417 Ingersoll Extension and the Graduate Center of the City University of New York, Brooklyn, NY 11210, USA.
7
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.
8
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.

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.

PMID:
24623816
PMCID:
PMC4027182
DOI:
10.1093/nar/gku175
[Indexed for MEDLINE]
Free PMC Article

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