title: Selection of intracellular single-domain antibodies targeting the HIV-1 Vpr protein by cytoplasmic yeast two-hybrid system.
abstract: The targeting of HIV-1 using antibodies is of high interest as molecular tools to better understand the biology of the virus or as a first step toward the design of new inhibitors targeting critical viral intracellular proteins. Small and highly stable llama-derived single-domain antibodies can often be functionally expressed as intracellular antibodies in the cytoplasm of eukaryotic cells. Using a selection method based on the Sos Recruitment System, a cytoplasmic yeast two-hybrid approach, we have isolated single-domain antibodies able to bind HIV-1 Vpr and Capside proteins in the yeast cytoplasm. One anti-Vpr single domain antibody was able to bind the HIV-1 regulatory Vpr protein in the cytoplasm of eukaryotic cells, leading to its delocalization from the nucleus to the cytoplasm. To our knowledge, this is the first description of a functional single-domain intrabody targeting HIV-1 Vpr, isolated using an in vivo cytoplasmic selection method that alleviates some limitations of the conventional yeast two-hybrid system.
authors.affiliation: Inserm U1068, CRCM, Marseille, France; Institut Paoli-Calmettes, Marseille, France; Aix-Marseille Université UM105, Marseille, France; CNRS, UMR7258, CRCM, Marseille, France.
authors.forename: Julie
authors.initials: J
authors.lastname: Matz
authors.affiliation: Institut Cochin, CNRS UMR8104, Paris, France; Université Paris Descartes, Paris, France; Inserm U1016, Paris, France.
authors.forename: Cécile
authors.initials: C
authors.lastname: Hérate
authors.affiliation: Institut Cochin, CNRS UMR8104, Paris, France; Université Paris Descartes, Paris, France; Inserm U1016, Paris, France.
authors.forename: Jérôme
authors.initials: J
authors.lastname: Bouchet
authors.affiliation: Inserm U1068, CRCM, Marseille, France; Institut Paoli-Calmettes, Marseille, France; Aix-Marseille Université UM105, Marseille, France; CNRS, UMR7258, CRCM, Marseille, France.
authors.forename: Nelson
authors.initials: N
authors.lastname: Dusetti
authors.affiliation: Inserm U1068, CRCM, Marseille, France; Institut Paoli-Calmettes, Marseille, France; Aix-Marseille Université UM105, Marseille, France; CNRS, UMR7258, CRCM, Marseille, France.
authors.forename: Odile
authors.initials: O
authors.lastname: Gayet
authors.affiliation: Inserm U1068, CRCM, Marseille, France; Institut Paoli-Calmettes, Marseille, France; Aix-Marseille Université UM105, Marseille, France; CNRS, UMR7258, CRCM, Marseille, France.
authors.forename: Daniel
authors.initials: D
authors.lastname: Baty
authors.affiliation: Institut Cochin, CNRS UMR8104, Paris, France; Université Paris Descartes, Paris, France; Inserm U1016, Paris, France.
authors.forename: Serge
authors.initials: S
authors.lastname: Benichou
authors.affiliation: Inserm U1068, CRCM, Marseille, France; Institut Paoli-Calmettes, Marseille, France; Aix-Marseille Université UM105, Marseille, France; CNRS, UMR7258, CRCM, Marseille, France.
authors.forename: Patrick
authors.initials: P
authors.lastname: Chames
mesh_terms: D002467:Cell Nucleus / Q000378:metabolism; D003593:Cytoplasm / Q000378:metabolism*; D015497:HIV-1 / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D011485:Protein Binding; D012441:Saccharomyces cerevisiae / Q000378:metabolism; D061905:Single-Domain Antibodies / Q000378:metabolism*; D014162:Transfection; D020798:Two-Hybrid System Techniques; D054325:vpr Gene Products, Human Immunodeficiency Virus / Q000276:immunology*
chemical_list: D061905:Single-Domain Antibodies; D054325:vpr Gene Products, Human Immunodeficiency Virus; C514912:vpr protein, Human immunodeficiency virus 1