title: Bimodal high-affinity association of Brd4 with murine leukemia virus integrase and mononucleosomes.
abstract: The importance of understanding the molecular mechanisms of murine leukemia virus (MLV) integration into host chromatin is highlighted by the development of MLV-based vectors for human gene-therapy. We have recently identified BET proteins (Brd2, 3 and 4) as the main cellular binding partners of MLV integrase (IN) and demonstrated their significance for effective MLV integration at transcription start sites. Here we show that recombinant Brd4, a representative of the three BET proteins, establishes complementary high-affinity interactions with MLV IN and mononucleosomes (MNs). Brd4(1-720) but not its N- or C-terminal fragments effectively stimulate MLV IN strand transfer activities in vitro. Mass spectrometry- and NMR-based approaches have enabled us to map key interacting interfaces between the C-terminal domain of BRD4 and the C-terminal tail of MLV IN. Additionally, the N-terminal fragment of Brd4 binds to both DNA and acetylated histone peptides, allowing it to bind tightly to MNs. Comparative analyses of the distributions of various histone marks along chromatin revealed significant positive correlations between H3- and H4-acetylated histones, BET protein-binding sites and MLV-integration sites. Our findings reveal a bimodal mechanism for BET protein-mediated MLV integration into select chromatin locations.
authors: Center for Retrovirus Research and College of Pharmacy, The Ohio State University, Columbus, Ohio 43210, USA, Department of Chemistry and Biochemistry, The Ohio State University, Columbus, Ohio 43210, Department of Microbiology, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA and Department of Pharmacology, Robert Wood Johnson Medical School, University of Medicine and Dentistry of New Jersey, Piscataway, NJ 08854, USA.
authors: Ross C
authors: RC
authors: Larue
authors: Matthew R
authors: MR
authors: Plumb
authors: Brandon L
authors: BL
authors: Crowe
authors: Nikoloz
authors: N
authors: Shkriabai
authors: Amit
authors: A
authors: Sharma
authors: Julia
authors: J
authors: DiFiore
authors: Nirav
authors: N
authors: Malani
authors: Sriram S
authors: SS
authors: Aiyer
authors: Monica J
authors: MJ
authors: Roth
authors: Frederic D
authors: FD
authors: Bushman
authors: Mark P
authors: MP
authors: Foster
authors: Mamuka
authors: M
authors: Kvaratskhelia
mesh_terms: D018797:Cell Cycle Proteins; D004247:DNA / Q000378:metabolism; D057809:HEK293 Cells; D006657:Histones / Q000378:metabolism; D006801:Humans; D019426:Integrases / Q000737:chemistry / Q000378:metabolism*; D009052:Leukemia Virus, Murine / Q000201:enzymology* / Q000502:physiology; D009687:Nuclear Proteins / Q000378:metabolism*; D009707:Nucleosomes / Q000378:metabolism*; D011485:Protein Binding; D054730:Protein Interaction Domains and Motifs; D014157:Transcription Factors / Q000378:metabolism*; D016662:Virus Integration; D000092468:Bromodomain Containing Proteins
chemical_list: D018797:Cell Cycle Proteins; D004247:DNA; D006657:Histones; D019426:Integrases; D009687:Nuclear Proteins; D009707:Nucleosomes; D014157:Transcription Factors; C441289:BRD4 protein, human; D000092468:Bromodomain Containing Proteins