In vitro processing of human immunodeficiency virus type 1 Gag virus-like particles.
Human immunodeficiency virus (HIV) Gag proteins are assembled into virus particles and then cleaved by the virion-associated HIV protease. Concomitant with Gag processing, doughnut-like HIV particles (the immature form) are converted to particles containing condensed cores (the mature form). Here we describe the in vitro processing of immature HIV Gag virus-like particles (VLP) by exogenously added HIV protease. Following delipidization, sequential processing of immature VLP showed that the matrix (MA)/capsid (CA) junction was cleaved faster than the CA/nucleocapsid (NC) junction, an altered order of processing when compared with authentic processing. When the in vitro processed VLP were analyzed on density gradients, most of the MA, CA-p15 intermediate, and NC were detected as a highly multimeric form, equivalent to the unprocessed VLP. In contrast, CA was found as a monomer dissociated from the multimeric CA-p15 following cleavage of the CA/NC junction. Electron microscopy revealed that the in vitro processing was accompanied by conversion of the doughnut-like particles to particles containing condensed cores and spherical outer shells. The cores, however, lacked core shells, which are normally observed for authentic HIV, suggesting that the in vitro processing of immature VLP failed to produce core shells.
Morikawa
Y
Y
The Kitasato Institute, Kitasato University, Shirokane 5-9-1, Minato-ku, Tokyo, 108-8642, Japan. ymorikawa@kitasato.or.jp
Shibuya
M
M
Goto
T
T
Sano
K
K
D000818:Animals; D004352:Drug Resistance, Microbial; D015683:Gene Products, gag / Q000378:metabolism* / Q000648:ultrastructure; D015497:HIV-1 / Q000187:drug effects / Q000378:metabolism* / Q000648:ultrastructure; D006801:Humans; D008051:Lipid Bilayers / Q000378:metabolism; D017830:Octoxynol / Q000494:pharmacology; D011499:Protein Processing, Post-Translational; D014771:Virion / Q000187:drug effects / Q000378:metabolism* / Q000648:ultrastructure
D015683:Gene Products, gag; D008051:Lipid Bilayers; D017830:Octoxynol