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Originally published In Press as doi:10.1074/jbc.M300248200 on March 14, 2003

J. Biol. Chem., Vol. 278, Issue 20, 18110-18116, May 16, 2003
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The Cationic Amphipathic alpha -Helix of HIV-1 Viral Protein R (Vpr) Binds to Nucleic Acids, Permeabilizes Membranes, and Efficiently Transfects Cells*

Emmanuel CoeytauxDagger , Dominique Coulaud§, Eric Le Cam§, Olivier DanosDagger , and Antoine KichlerDagger

From Dagger  Généthon-Unité Mixte de Recherche 8115 CNRS, 1 bis rue de l'Internationale, BP 60, F-91002 Evry and § Interactions Moléculaires et Cancer, UMR 8126 CNRS, Institut Gustave Roussy, 94805 Villejuif, France

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.

* This work was supported by the Association Française contre les Myopathies.The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

To whom correspondence should be addressed. Tel.: 33-1-69-47-10-28; Fax: 33-1-60-77-86-98; E-mail: akich@genethon.fr.

Copyright © 2003 by The American Society for Biochemistry and Molecular Biology, Inc.
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