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Journal of Virology, July 2005, p. 8629-8636, Vol. 79, No. 13
0022-538X/05/$08.00+0     doi:10.1128/JVI.79.13.8629-8636.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Detection of Human Immunodeficiency Virus Type 1 Nef and CD4 Physical Interaction in Living Human Cells by Using Bioluminescence Resonance Energy Transfer

David Cluet,1 Christophe Bertsch,{dagger} Christian Beyer,1 Liliane Gloeckler,1 Mathieu Erhardt,1,{ddagger} Jean-Pierre Gut,1 Jean-Luc Galzi,2 and Anne-Marie Aubertin1*

INSERM-ULP U544, Université Louis Pasteur, Institut de Virologie 3 Rue Koeberlé, 67000 Strasbourg, France,1 CNRS UPR 9050, Département Récepteurs et Protéines membranaires, ESBS, Boulevard Sébastien Brandt, 67400 Illkirch, France2

Received 20 January 2005/ Accepted 21 February 2005

CD4 down-regulation by human immunodeficiency virus type 1 (HIV-1) Nef protein is a key function for virus virulence. This activity may be mediated by a direct Nef-CD4 interaction. We investigated the formation, in situ, of such a complex between proteins using bioluminescence resonance energy transfer technology and coimmunoprecipitations. Our data clearly demonstrate that Nef and CD4 interact in intact human cells. Moreover, our results clearly indicate that the dileucine motif of the CD4 cytoplasmic domain, critical for the Nef-induced CD4 down-regulation, is not implicated in the Nef/CD4 complex formation in the cellular context.

* Corresponding author. Mailing address: INSERM-ULP U544, Université Louis Pasteur, Institut de Virologie, 3 Rue Koeberlé, 67000 Strasbourg, France. Phone: (33) 3 90 24 37 44. Fax: (33) 3 90 24 37 23. E-mail: An.Aubertin{at}viro-ulp.u-strasbg.fr.

{dagger} Present address: Laboratoire Vigne Biotechnologies et Environnement-LVBE- 33, rue de Herrlisheim, 68000 Colmar, France.

{ddagger} Present address: IBMP, 12 Rue du Général Zimmer, 67084 Strasbourg Cedex, France.

Journal of Virology, July 2005, p. 8629-8636, Vol. 79, No. 13
0022-538X/05/$08.00+0     doi:10.1128/JVI.79.13.8629-8636.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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