The Cationic Amphipathic α-Helix of HIV-1 Viral Protein R (Vpr) Binds to Nucleic Acids, Permeabilizes Membranes, and Efficiently Transfects Cells
| Autor: | Eric Le Cam, Dominique Coulaud, Antoine Kichler, Olivier Danos, Emmanuel Coeytaux |
|---|---|
| Přispěvatelé: | Laboratoire de Biologie et de Pharmacologie Appliquée (LBPA), École normale supérieure - Cachan (ENS Cachan)-Centre National de la Recherche Scientifique (CNRS), Institut Gustave Roussy (IGR), Signalisation, noyaux et innovations en cancérologie (UMR8126), Université Paris-Sud - Paris 11 (UP11)-Institut Gustave Roussy (IGR)-Centre National de la Recherche Scientifique (CNRS), Centre de recherche et d'applications sur les thérapies géniques (CRATG), Université d'Évry-Val-d'Essonne (UEVE)-Centre National de la Recherche Scientifique (CNRS)-Généthon, Généthon, Centre National de la Recherche Scientifique (CNRS)-Institut Gustave Roussy (IGR)-Université Paris-Sud - Paris 11 (UP11), Université d'Évry-Val-d'Essonne (UEVE)-Généthon-Centre National de la Recherche Scientifique (CNRS) |
| Jazyk: | angličtina |
| Rok vydání: | 2003 |
| Předmět: |
Erythrocytes
viruses [SDV]Life Sciences [q-bio] Endocytic cycle Plasma protein binding Biology Transfection Biochemistry Cell Line Cell membrane 03 medical and health sciences chemistry.chemical_compound Plasmid Protein structure Ethidium Nucleic Acids medicine Humans Polylysine Transgenes Molecular Biology Cells Cultured ComputingMilieux_MISCELLANEOUS 030304 developmental biology 0303 health sciences Gene Products vpr 030302 biochemistry & molecular biology Cell Membrane Cell Biology DNA Flow Cytometry Endocytosis Intercalating Agents 3. Good health Protein Structure Tertiary Microscopy Electron medicine.anatomical_structure chemistry Nucleic acid Peptides Plasmids Protein Binding |
| Zdroj: | Journal of Biological Chemistry Journal of Biological Chemistry, American Society for Biochemistry and Molecular Biology, 2003, 278 (20), pp.18110-18116. ⟨10.1074/jbc.m300248200⟩ Journal of Biological Chemistry, 2003, 278 (20), pp.18110-18116. ⟨10.1074/jbc.m300248200⟩ |
| ISSN: | 0021-9258 1083-351X |
| DOI: | 10.1074/jbc.m300248200⟩ |
| Popis: | Viral protein R (Vpr) is a small protein of 96 amino acids that is conserved among the lentiviruses human immunodeficiency virus type 1 (HIV-1), HIV-2, and simian immunodeficiency virus. We recently sought to determine whether the karyophilic properties of Vpr, as well as its ability to bind nucleic acids, could be used to deliver DNA into cells. We have found that the C-terminal domain of Vpr-(52-96) is able to efficiently transfect various cell lines. Here, we show that the shortest active sequence for gene transfer corresponds to the domain that adopts a alpha-helix conformation. DNA binding studies and permeabilization assays performed on cells demonstrated that the peptides that are efficient in transfection condense plasmid DNA and are membranolytic. Electron microscopy studies and transfection experiments performed in the presence of inhibitors of the endocytic processes indicated that the major entry pathway of Vpr-DNA complexes is through endocytosis. Taken together, the results show that the cationic C-terminal alpha-helix of Vpr has DNA-condensing as well as membrane-destabilizing capabilities, both properties that are indispensable for efficient DNA transfection. |
| Databáze: | OpenAIRE |
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