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Journal of Virology, June 2007, p. 6434-6445, Vol. 81, No. 12
0022-538X/07/$08.00+0     doi:10.1128/JVI.02757-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Electrostatic Interactions Drive Membrane Association of the Human Immunodeficiency Virus Type 1 Gag MA Domain

Amanda K. Dalton,¹ Danso Ako-Adjei,^1, Paul S. Murray,^2, Diana Murray,² and Volker M. Vogt^1*

Department of Molecular Biology and Genetics, Biotechnology Building, Cornell University, Ithaca, New York 14853,¹ Department of Microbiology and Immunology and Institute for Computational Biomedicine, Weill Cornell Medical College, New York, New York 10021²

Received 14 December 2006/ Accepted 20 March 2007

The assembly of most retroviruses occurs at the plasma membrane. Membrane association is directed by MA, the N-terminal domain of the Gag structural protein. For human immunodeficiency virus type 1 (HIV-1), this association is mediated in part by a myristate fatty acid modification. Conflicting evidence has been presented on the relative importance of myristoylation, of ionic interactions between protein and membrane, and of Gag multimerization in membrane association in vivo. We addressed these questions biochemically by determining the affinity of purified myristoylated HIV-1 MA for liposomes of defined composition, both for monomeric and for dimeric forms of the protein. Myristoylation increases the barely detectable intrinsic affinity of the apo-protein for liposomes by only 10-fold, and the resulting affinity is still weak, similar to that of the naturally nonmyristoylated MA of Rous sarcoma virus. Membrane binding of HIV-1 MA is absolutely dependent on the presence of negatively charged lipid and is abrogated at high ionic strength. Forced dimerization of MA increases its membrane affinity by several orders of magnitude. When green fluorescent protein fusions of monomeric or dimeric MA are expressed in cells, the dimeric but not the monomeric protein becomes strongly membrane associated. Computational modeling supports these results and suggests a molecular mechanism for the modest effect of myristoylation on binding, wherein the membrane provides a hydrophobic environment for the myristate that is energetically similar to that provided by the protein. Overall, the results imply that the driving force for membrane association stems largely from ionic interactions between multimerized Gag and negatively charged phospholipids.

Published ahead of print on 28 March 2007.

D.A.-A. and P.S.M. contributed equally to this study.

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