Efficient DNA transfection mediated by the C-terminal domain of human immunodeficiency virus type 1 viral protein R.
Viral protein R (Vpr) of human immunodeficiency virus type 1 is produced late in the virus life cycle and is assembled into the virion through binding to the Gag protein. It is known to play a significant role early in the viral life cycle by facilitating the nuclear import of the preintegration complex in nondividing cells. Vpr is also able to interact with nucleic acids, and we show here that it induces condensation of plasmid DNA. We have explored the possibility of using these properties in DNA transfection experiments. We report that the C-terminal half of the protein (Vpr(52-96)) mediates DNA transfection in a variety of human and nonhuman cell lines with efficiencies comparable to those of the best-known transfection agents. Compared with polylysine, a standard polycationic transfection reagent, Vpr(52-96) was 10- to 1,000-fold more active. Vpr(52-96)-DNA complexes were able to reach the cell nucleus through a pH-independent mechanism. These observations possibly identify an alternate pathway for DNA transfection.
Kichler
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Généthon III, CNRS URA 1923, Evry, France. akich@genethon.fr
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Leborgne
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Druillennec
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Lenoir
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Coulaud
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Delain
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Le Cam
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Roques
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Danos
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D000595:Amino Acid Sequence; D000818:Animals; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D002453:Cell Cycle; D002460:Cell Line; D002467:Cell Nucleus / Q000235:genetics / Q000378:metabolism; D011232:Chemical Precipitation; D002738:Chloroquine / Q000494:pharmacology; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism / Q000648:ultrastructure; D016336:Gene Products, vpr / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D017930:Genes, Reporter / Q000235:genetics; D015497:HIV-1; D006801:Humans; D006863:Hydrogen-Ion Concentration; D018942:Macrolides; D051379:Mice; D008854:Microscopy, Electron; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D010957:Plasmids / Q000235:genetics / Q000378:metabolism; D011107:Polylysine / Q000378:metabolism; D011401:Promoter Regions, Genetic / Q000235:genetics; D011485:Protein Binding; D015533:Transcriptional Activation / Q000187:drug effects / Q000235:genetics; D014162:Transfection / Q000379:methods*; D054325:vpr Gene Products, Human Immunodeficiency Virus
D000900:Anti-Bacterial Agents; D004268:DNA-Binding Proteins; D016336:Gene Products, vpr; D018942:Macrolides; D010446:Peptide Fragments; D054325:vpr Gene Products, Human Immunodeficiency Virus; D011107:Polylysine; D002738:Chloroquine; C040929:bafilomycin A1