1. Live-Cell Imaging Reveals Multiple Interactions between Epstein-Barr Virus Nuclear Antigen 1 and Cellular Chromatin during Interphase and Mitosis
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Maïté Coppey-Moisan, Tristan Piolot, Vincent Maréchal, Gabriel Dos Reis, Christophe Klein, Nathalie Jourdan, Marc Tramier, Frédérique Quignon, Aude Jobart-Malfait, Adaptation Biologique et Vieillissement = Biological Adaptation and Ageing (B2A), Institut National de la Santé et de la Recherche Médicale (INSERM)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-Institut de Biologie Paris Seine (IBPS), Institut National de la Santé et de la Recherche Médicale (INSERM)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS), Institut Jacques Monod (IJM (UMR_7592)), Université Paris Diderot - Paris 7 (UPD7)-Centre National de la Recherche Scientifique (CNRS), Centre de Recherche des Cordeliers (CRC (UMR_S 872)), Université Pierre et Marie Curie - Paris 6 (UPMC)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Institut de Génétique et Développement de Rennes (IGDR), Université de Rennes (UR)-Centre National de la Recherche Scientifique (CNRS)-Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique ), Centre de Recherche des Cordeliers ( CRC (UMR_S 872) ), Université Pierre et Marie Curie - Paris 6 ( UPMC ) -Université Paris Descartes - Paris 5 ( UPD5 ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Centre National de la Recherche Scientifique ( CNRS ), Institut Jacques Monod ( IJM ), Université Paris Diderot - Paris 7 ( UPD7 ) -Centre National de la Recherche Scientifique ( CNRS ), Institut de Génétique et Développement de Rennes ( IGDR ), Université de Rennes 1 ( UR1 ), Université de Rennes ( UNIV-RENNES ) -Université de Rennes ( UNIV-RENNES ) -Centre National de la Recherche Scientifique ( CNRS ) -Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique ), UPMC, INSERM, Université Paris Descartes - Paris 5 (UPD5)-Université Pierre et Marie Curie - Paris 6 (UPMC)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Université de Rennes 1 (UR1), Université de Rennes (UNIV-RENNES)-Université de Rennes (UNIV-RENNES)-Centre National de la Recherche Scientifique (CNRS)-Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique ), and De Villemeur, Hervé
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MESH : Cell Line ,viruses ,[SDV]Life Sciences [q-bio] ,Gene Expression ,MESH : Hepatocytes ,MESH: Hepatocytes ,Chromatin bridge ,0302 clinical medicine ,MESH : Protein Transport ,hemic and lymphatic diseases ,MESH : Protein Stability ,0303 health sciences ,MESH : Chromatin ,MESH : Epstein-Barr Virus Nuclear Antigens ,MESH : Interphase ,Protein Stability ,RNA-Binding Proteins ,MESH : Protein Binding ,Cell cycle ,MESH : Mitosis ,Chromatin ,Virus-Cell Interactions ,Protein Transport ,030220 oncology & carcinogenesis ,Premature chromosome condensation ,Interphase ,MESH: Cell Nucleolus ,MESH : Carrier Proteins ,MESH: Epstein-Barr Virus Nuclear Antigens ,Cell Nucleolus ,Plasmids ,Protein Binding ,MESH: Protein Transport ,MESH: Gene Expression ,Immunology ,Mitosis ,MESH: Carrier Proteins ,[SDV.BC]Life Sciences [q-bio]/Cellular Biology ,Biology ,Microbiology ,Cell Line ,MESH: Chromatin ,MESH: Interphase ,03 medical and health sciences ,G2 phase ,Prophase ,MESH: Plasmids ,MESH: Protein Stability ,Virology ,MESH: HMGB2 Protein ,HMGB2 Protein ,Humans ,MESH: Protein Binding ,[SDV.BC] Life Sciences [q-bio]/Cellular Biology ,030304 developmental biology ,MESH: Humans ,[ SDV.BC ] Life Sciences [q-bio]/Cellular Biology ,MESH : Humans ,MESH: Mitosis ,MESH : HMGB2 Protein ,Molecular biology ,MESH: Cell Line ,MESH : Gene Expression ,Epstein-Barr Virus Nuclear Antigens ,MESH : Plasmids ,Insect Science ,MESH : Cell Nucleolus ,Hepatocytes ,Carrier Proteins - Abstract
Epstein-Barr virus (EBV) establishes a life-long latent infection in humans. In proliferating latently infected cells, EBV genomes persist as multiple episomes that undergo one DNA replication event per cell cycle and remain attached to the mitotic chromosomes. EBV nuclear antigen 1 (EBNA-1) binding to the episome and cellular genome is essential to ensure proper episome replication and segregation. However, the nature and regulation of EBNA-1 interaction with chromatin has not been clearly elucidated. This activity has been suggested to involve EBNA-1 binding to DNA, duplex RNA, and/or proteins. EBNA-1 binding protein 2 (EBP2), a nucleolar protein, has been proposed to act as a docking protein for EBNA-1 on mitotic chromosomes. However, there is no direct evidence thus far for EBP2 being associated with EBNA-1 during mitosis. By combining video microscopy and Förster resonance energy transfer (FRET) microscopy, we demonstrate here for the first time that EBNA-1 and EBP2 interact in the nucleoplasm, as well as in the nucleoli during interphase. However, in strong contrast to the current proposed model, we were unable to observe any interaction between EBNA-1 and EBP2 on mitotic chromosomes. We also performed a yeast double-hybrid screening, followed by a FRET analysis, that led us to identify HMGB2 (high-mobility group box 2), a well-known chromatin component, as a new partner for EBNA-1 on chromatin during interphase and mitosis. Although the depletion of HMGB2 partly altered EBNA-1 association with chromatin in HeLa cells during interphase and mitosis, it did not significantly impact the maintenance of EBV episomes in Raji cells.
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- 2012