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J. Biol. Chem., Vol. 278, Issue 37, 35211-35219, September 12, 2003

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Active cAMP-dependent Protein Kinase Incorporated within Highly Purified HIV-1 Particles Is Required for Viral Infectivity and Interacts with Viral Capsid Protein^*

Christine Cartier , Bénédicte Hemonnot , Bernard Gay, Martine Bardy, Céline Sanchiz, Christian Devaux and Laurence Briant ¶

From the Laboratoire Infections Rétrovirales et Signalisation Cellulaire, Centre National de la Recherche Scientifique, UMR 5121-UM1, Institut de Biologie, CS 89508, 34960 Montpellier Cedex 2, France

Host cell components, including protein kinases such as ERK-2/mitogen-activated protein kinase, incorporated within human immunodeficiency virus type 1 (HIV-1) virions play a pivotal role in the ability of HIV to infect and replicate in permissive cells. The present work provides evidence that the catalytic subunit of cAMP-dependent protein kinase (C-PKA) is packaged within HIV-1 virions as demonstrated using purified subtilisin-digested viral particles. Virus-associated C-PKA was shown to be enzymatically active and able to phosphorylate synthetic substrate in vitro. Suppression of virion-associated C-PKA activity by specific synthetic inhibitor had no apparent effect on viral precursor maturation and virus assembly. However, virus-associated C-PKA activity was demonstrated to regulate HIV-1 infectivity as assessed by single round infection assays performed by using viruses produced from cells expressing an inactive form of C-PKA. In addition, virus-associated C-PKA was found to co-precipitate with and to phosphorylate the CAp24^gag protein. Altogether our results indicate that virus-associated C-PKA regulates HIV-1 infectivity, possibly by catalyzing phosphorylation of the viral CAp24^gag protein.

Received for publication, February 5, 2003 , and in revised form, June 10, 2003.

^* This work was supported by institutional funds from CNRS, the French agency against AIDS (ANRS), and the Foundation pour la Recherche Médicale-Sidaction program "Ensemble contre le SIDA." 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.

Recipient of a grant from Ensemble contre le SIDA.

Supported by the ANRS.

^¶ To whom correspondence should be addressed: CNRS UMR 5121-UM1, Institut de Biologie, 4 Bd Henri IV, 34960 Montpellier, France. Tel.: 33-4-67-60-86-60; Fax: 33-4-67-60-44-20; E-mail: laurence.briant{at}univ-montp1.fr.

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