title: Manipulation of the N-terminal sequence of the Borna disease virus X protein improves its mitochondrial targeting and neuroprotective potential.
abstract: To favor their replication, viruses express proteins that target diverse mammalian cellular pathways. Due to the limited size of many viral genomes, such proteins are endowed with multiple functions, which require targeting to different subcellular compartments. One salient example is the X protein of Borna disease virus, which is expressed both at the mitochondria and in the nucleus. Moreover, we recently demonstrated that mitochondrial X protein is neuroprotective. In this study, we sought to examine the mechanisms whereby the X protein transits between subcellular compartments and to define its localization signals, to enhance its mitochondrial accumulation and thus, potentially, its neuroprotective activity. We transfected plasmids expressing fusion proteins bearing different domains of X fused to enhanced green fluorescent protein (eGFP) and compared their subcellular localization to that of eGFP. We observed that the 5-16 domain of X was responsible for both nuclear export and mitochondrial targeting and identified critical residues for mitochondrial localization. We next took advantage of these findings and constructed mutant X proteins that were targeted only to the mitochondria. Such mutants exhibited enhanced neuroprotective properties in compartmented cultures of neurons grown in microfluidic chambers, thereby confirming the parallel between mitochondrial accumulation of the X protein and its neuroprotective potential.-Ferré C. A., Davezac, N., Thouard, A., Peyrin, J. M., Belenguer, P., Miquel, M.-C., Gonzalez-Dunia, D., Szelechowski, M. Manipulation of the N-terminal sequence of the Borna disease virus X protein improves its mitochondrial targeting and neuroprotective potential.
authors.affiliation: *INSERM, Unité Mixte de Recherche (UMR) 1043, Centre de Physiopathologie de Toulouse Purpan (CPTP), Toulouse, France; Centre National de la Recherche Scientifique (CNRS), UMR 5282, Toulouse, France; Université Toulouse III Paul Sabatier, Toulouse, France; CNRS UMR 5547, Centre de Biologie du Développement, Toulouse, France; CNRS UMR 8256, Biological Adaptation and Aging, Institut de Biologie Paris Seine, Université Pierre et Marie Curie, Paris, France; and Université Pierre et Marie Curie, Sorbonne Universités, Paris, France.
authors.forename: Cécile A
authors.initials: CA
authors.lastname: Ferré
authors.affiliation: *INSERM, Unité Mixte de Recherche (UMR) 1043, Centre de Physiopathologie de Toulouse Purpan (CPTP), Toulouse, France; Centre National de la Recherche Scientifique (CNRS), UMR 5282, Toulouse, France; Université Toulouse III Paul Sabatier, Toulouse, France; CNRS UMR 5547, Centre de Biologie du Développement, Toulouse, France; CNRS UMR 8256, Biological Adaptation and Aging, Institut de Biologie Paris Seine, Université Pierre et Marie Curie, Paris, France; and Université Pierre et Marie Curie, Sorbonne Universités, Paris, France.
authors.forename: Noélie
authors.initials: N
authors.lastname: Davezac
authors.affiliation: *INSERM, Unité Mixte de Recherche (UMR) 1043, Centre de Physiopathologie de Toulouse Purpan (CPTP), Toulouse, France; Centre National de la Recherche Scientifique (CNRS), UMR 5282, Toulouse, France; Université Toulouse III Paul Sabatier, Toulouse, France; CNRS UMR 5547, Centre de Biologie du Développement, Toulouse, France; CNRS UMR 8256, Biological Adaptation and Aging, Institut de Biologie Paris Seine, Université Pierre et Marie Curie, Paris, France; and Université Pierre et Marie Curie, Sorbonne Universités, Paris, France.
authors.forename: Anne
authors.initials: A
authors.lastname: Thouard
authors.affiliation: *INSERM, Unité Mixte de Recherche (UMR) 1043, Centre de Physiopathologie de Toulouse Purpan (CPTP), Toulouse, France; Centre National de la Recherche Scientifique (CNRS), UMR 5282, Toulouse, France; Université Toulouse III Paul Sabatier, Toulouse, France; CNRS UMR 5547, Centre de Biologie du Développement, Toulouse, France; CNRS UMR 8256, Biological Adaptation and Aging, Institut de Biologie Paris Seine, Université Pierre et Marie Curie, Paris, France; and Université Pierre et Marie Curie, Sorbonne Universités, Paris, France.
authors.forename: Jean-Michel
authors.initials: JM
authors.lastname: Peyrin
authors.affiliation: *INSERM, Unité Mixte de Recherche (UMR) 1043, Centre de Physiopathologie de Toulouse Purpan (CPTP), Toulouse, France; Centre National de la Recherche Scientifique (CNRS), UMR 5282, Toulouse, France; Université Toulouse III Paul Sabatier, Toulouse, France; CNRS UMR 5547, Centre de Biologie du Développement, Toulouse, France; CNRS UMR 8256, Biological Adaptation and Aging, Institut de Biologie Paris Seine, Université Pierre et Marie Curie, Paris, France; and Université Pierre et Marie Curie, Sorbonne Universités, Paris, France.
authors.forename: Pascale
authors.initials: P
authors.lastname: Belenguer
authors.affiliation: *INSERM, Unité Mixte de Recherche (UMR) 1043, Centre de Physiopathologie de Toulouse Purpan (CPTP), Toulouse, France; Centre National de la Recherche Scientifique (CNRS), UMR 5282, Toulouse, France; Université Toulouse III Paul Sabatier, Toulouse, France; CNRS UMR 5547, Centre de Biologie du Développement, Toulouse, France; CNRS UMR 8256, Biological Adaptation and Aging, Institut de Biologie Paris Seine, Université Pierre et Marie Curie, Paris, France; and Université Pierre et Marie Curie, Sorbonne Universités, Paris, France.
authors.forename: Marie-Christine
authors.initials: MC
authors.lastname: Miquel
authors.affiliation: *INSERM, Unité Mixte de Recherche (UMR) 1043, Centre de Physiopathologie de Toulouse Purpan (CPTP), Toulouse, France; Centre National de la Recherche Scientifique (CNRS), UMR 5282, Toulouse, France; Université Toulouse III Paul Sabatier, Toulouse, France; CNRS UMR 5547, Centre de Biologie du Développement, Toulouse, France; CNRS UMR 8256, Biological Adaptation and Aging, Institut de Biologie Paris Seine, Université Pierre et Marie Curie, Paris, France; and Université Pierre et Marie Curie, Sorbonne Universités, Paris, France daniel.dunia@inserm.fr mszelechowski@gmail.com.
authors.forename: Daniel
authors.initials: D
authors.lastname: Gonzalez-Dunia
authors.affiliation: *INSERM, Unité Mixte de Recherche (UMR) 1043, Centre de Physiopathologie de Toulouse Purpan (CPTP), Toulouse, France; Centre National de la Recherche Scientifique (CNRS), UMR 5282, Toulouse, France; Université Toulouse III Paul Sabatier, Toulouse, France; CNRS UMR 5547, Centre de Biologie du Développement, Toulouse, France; CNRS UMR 8256, Biological Adaptation and Aging, Institut de Biologie Paris Seine, Université Pierre et Marie Curie, Paris, France; and Université Pierre et Marie Curie, Sorbonne Universités, Paris, France daniel.dunia@inserm.fr mszelechowski@gmail.com.
authors.forename: Marion
authors.initials: M
authors.lastname: Szelechowski
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001224:Aspartic Acid / Q000235:genetics / Q000378:metabolism; D001369:Axons / Q000187:drug effects / Q000378:metabolism; D015153:Blotting, Western; D001891:Borna disease virus / Q000235:genetics* / Q000378:metabolism; D019556:COS Cells; D002478:Cells, Cultured; D002522:Chlorocebus aethiops; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D008856:Microscopy, Fluorescence; D008928:Mitochondria / Q000378:metabolism*; D008969:Molecular Sequence Data; D009154:Mutation; D009474:Neurons / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D019913:Nuclear Localization Signals / Q000235:genetics; D017386:Sequence Homology, Amino Acid; D014764:Viral Proteins / Q000235:genetics* / Q000378:metabolism
chemical_list: D019913:Nuclear Localization Signals; D014764:Viral Proteins; D049452:Green Fluorescent Proteins; D001224:Aspartic Acid
keywords: neuroprotection; nuclear export; subcellular addressing