Electrostatic interactions drive membrane association of the human immunodeficiency virus type 1 Gag MA domain.
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.
Dalton
Amanda K
AK
Department of Molecular Biology and Genetics, Biotechnology Building, Cornell University, Ithaca, NY 14853, USA.
Ako-Adjei
Danso
D
Murray
Paul S
PS
Murray
Diana
D
Vogt
Volker M
VM
D000818:Animals; D002462:Cell Membrane / Q000378:metabolism*; D002645:Chickens; D019281:Dimerization; D004926:Escherichia coli / Q000378:metabolism; D005227:Fatty Acids / Q000737:chemistry; D015683:Gene Products, gag / Q000737:chemistry*; D015497:HIV-1 / Q000378:metabolism*; D006801:Humans; D007700:Kinetics; D019814:Myristic Acid / Q000737:chemistry; D010743:Phospholipids / Q000737:chemistry; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D055672:Static Electricity
D005227:Fatty Acids; D015683:Gene Products, gag; D010743:Phospholipids; D019814:Myristic Acid