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J. Biol. Chem., Vol. 282, Issue 48, 35163-35168, November 30, 2007

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CD4 Interacts Constitutively with Multiple CCR5 at the Plasma Membrane of Living Cells

A FLUORESCENCE RECOVERY AFTER PHOTOBLEACHING AT VARIABLE RADII APPROACH^*

Aurélie-Marie Baker¹, Aude Saulière², Gérald Gaibelet³, Bernard Lagane, Serge Mazères, Marie Fourage, Françoise Bachelerie, Laurence Salomé, André Lopez⁴, and Fabrice Dumas⁵

From the Institut de Pharmacologie et Biologie Structurale, UMR CNRS-Université Paul Sabatier 5089, 205 Route de Narbonne, 31062 Toulouse cedex, France, Unité de Pathogénie Virale Moléculaire, Institut Pasteur, INSERM U819, 75015 Paris, France

The entry of human immunodeficiency virus into target cells requires successive interactions of the viral envelope glycoprotein gp120 with CD4 and the chemokine receptors CCR5 or CXCR4. We previously demonstrated, by Förster resonance energy transfer experiments, the constitutive association of CD4 and CCR5 at the surface of living cells. We therefore speculated that this interaction may correlate with compartmentalization of CD4 and CCR5 within the plasma membrane. Here, we characterize the lateral distribution, the dynamics, and the stoichiometry of these receptors in living cells stably expressing CD4 and/or CCR5 by means of fluorescence recovery after photobleaching at variable radii experiments. We found that (i) these receptors expressed alone are confined into 1-µm-sized domains, (ii) CD4-CCR5 associations occur outside and inside smaller domains, and (iii) these interactions involve multiple CCR5 molecules per CD4.

Received for publication, July 9, 2007 , and in revised form, August 22, 2007.

^* This work has been supported by Agence Nationale de Recherche sur le SIDA (ANRS) and CNRS. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

¹ Supported by Fonds Social Européen.

² Supported by Ministère de l'Enseignement Supérieur et de la Recherche.

³ Supported by ANRS.

⁴ To whom correspondence may be addressed. Tel.: 561175468; Fax: 561175994; E-mail: Andre.Lopez{at}ipbs.fr.

⁵ To whom correspondence may be addressed. Tel.: 561175468; Fax: 561175994; E-mail: Fabrice.Dumas{at}ipbs.fr.

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