Host ABCE1 is at plasma membrane HIV assembly sites and its dissociation from Gag is linked to subsequent events of virus production.
In primate cells, assembly of a single HIV-1 capsid involves multimerization of thousands of Gag polypeptides, typically at the plasma membrane. Although studies support a model in which HIV-1 assembly proceeds through complexes containing Gag and the cellular adenosine triphosphatase ABCE1 (also termed HP68 or ribonuclease L inhibitor), whether these complexes constitute true assembly intermediates remains controversial. Here we demonstrate by pulse labeling in primate cells that a population of Gag associates with endogenous ABCE1 within minutes of translation. In the next approximately 2 h, Gag-ABCE1 complexes increase in size to approximately that of immature capsids. Dissociation of ABCE1 from Gag correlates closely with Gag processing during virion maturation and occurs much less efficiently when the HIV-1 protease is inactivated. Finally, quantitative double-label immunogold electron microscopy reveals that ABCE1 is recruited to sites of assembling wild-type Gag at the plasma membrane but not to sites of an assembly-defective Gag mutant at the plasma membrane. Together these findings demonstrate that a population of Gag present at plasma membrane sites of assembly associates with ABCE1 throughout capsid formation until the onset of virus maturation, which is then followed by virus release. Moreover, the data suggest a linkage between Gag-ABCE1 dissociation and subsequent events of virion production.
Dooher
Julia E
JE
Department of Pathobiology, University of Washington, 1959 NE Pacific Street, Seattle, WA 98195, USA.
Schneider
Bobbie L
BL
Reed
Jonathan C
JC
Lingappa
Jaisri R
JR
D018528:ATP-Binding Cassette Transporters / Q000378:metabolism*; D000818:Animals; D019556:COS Cells; D002213:Capsid / Q000378:metabolism; D002462:Cell Membrane / Q000378:metabolism*; D002522:Chlorocebus aethiops; D015683:Gene Products, gag / Q000378:metabolism*; D015750:Genes, gag; D016655:HIV Core Protein p24 / Q000378:metabolism; D015497:HIV-1 / Q000737:chemistry* / Q000378:metabolism; D007700:Kinetics; D009154:Mutation; D011498:Protein Precursors / Q000378:metabolism; D019065:Virus Assembly
D018528:ATP-Binding Cassette Transporters; D015683:Gene Products, gag; D016655:HIV Core Protein p24; D011498:Protein Precursors; C059767:p55 gag precursor protein, Human immunodeficiency virus 1