Ku80 participates in the targeting of retroviral transgenes to the chromatin of CHO cells
| Autor: | Christel Masson, Stéphanie Bury-Moné, Jean-François Mouscadet, Corinne Brachet-Ducos, Olivier Delelis, Asier Sáez-Cirión, Laurence Jeanson-Leh, Damien Schoëvaërt-Brossault, Frédéric Subra, Elvire Guiot |
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| Přispěvatelé: | Laboratoire de Biologie et de Pharmacologie Appliquée (LBPA), École normale supérieure - Cachan (ENS Cachan)-Centre National de la Recherche Scientifique (CNRS), Régulation des Infections Rétrovirales, Institut Pasteur [Paris] (IP), Laboratoire d'Andrologie, AP-HP Hôpital Bicêtre (Le Kremlin-Bicêtre), Laboratoire d'Analyse d'Images en Pathologie Cellulaire, Université Paris Diderot - Paris 7 (UPD7), Institut Pasteur [Paris] |
| Jazyk: | angličtina |
| Rok vydání: | 2007 |
| Předmět: |
Ku80
Transgene Virus Integration Immunology Genetic Vectors MESH: Cricetinae MESH: Transgenes CHO Cells [SDV.BC]Life Sciences [q-bio]/Cellular Biology Biology Microbiology MESH: Chromatin 03 medical and health sciences Transduction (genetics) Cricetulus MESH: Cricetulus Transcription (biology) MESH: CHO Cells MESH: Genetic Vectors Virology Cricetinae Animals Humans MESH: Animals Transgenes MESH: In Situ Hybridization Fluorescence Promoter Regions Genetic Ku Autoantigen MESH: Antigens Nuclear In Situ Hybridization Fluorescence 030304 developmental biology 0303 health sciences Ku70 MESH: Humans 030302 biochemistry & molecular biology Antigens Nuclear Provirus Molecular biology Chromatin Genome Replication and Regulation of Viral Gene Expression DNA-Binding Proteins MESH: Promoter Regions (Genetics) Retroviridae MESH: Retroviridae Insect Science MESH: Virus Integration MESH: DNA-Binding Proteins |
| Zdroj: | Journal of Virology Journal of Virology, 2007, 81 (15), pp.7924-32. ⟨10.1128/JVI.02015-06⟩ Journal of Virology, American Society for Microbiology, 2007, 81 (15), pp.7924-32. ⟨10.1128/JVI.02015-06⟩ |
| ISSN: | 0022-538X 1098-5514 |
| DOI: | 10.1128/JVI.02015-06⟩ |
| Popis: | Free in PubMed Central : http://www.pubmedcentral.nih.gov/picrender.fcgi?tool=pmcentrez&artid=1951289&blobtype=pdf; The heterodimer Ku70/80 Ku is the DNA-binding component of the DNA-PK complex required for the nonhomologous end-joining pathway. It participates in numerous nuclear processes, including telomere and chromatin structure maintenance, replication, and transcription. Ku interacts with retroviral preintegration complexes and is thought to interfere with the retroviral replication cycle, in particular the formation of 2-long terminal repeat (LTR) viral DNA circles, viral DNA integration, and transcription. We describe here the effect of Ku80 on both provirus integration and the resulting transgene expression in cells transduced with retroviral vectors. We found that transgene expression was systematically higher in Ku80-deficient xrs6 cells than in Ku80-expressing CHO cells. This higher expression was observed irrespective of the presence of the viral LTR and was also not related to the nature of the promoter. Real-time PCR monitoring of the early viral replicative steps demonstrated that the absence of Ku80 does not affect the efficiency of transduction. We analyzed the transgene distributions localization in nucleus by applying a three-dimensional reconstruction model to two-dimensional fluorescence in situ hybridization images. This indicated that the presence of Ku80 resulted in a bias toward the transgenes being located at the periphery of the nucleus associated with their being repressed; in the absence of this factor the transgenes tend to be randomly distributed and actively expressed. Therefore, although not strictly required for retroviral integration, Ku may be involved in targeting retroviral elements to chromatin domains prone to gene silencing. |
| Databáze: | OpenAIRE |
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