Complete inhibition of human immunodeficiency virus Gag myristoylation is necessary for inhibition of particle budding.
Myristoylation of human immunodeficiency virus (HIV) Gag protein is essential for virus particle budding. Two reactions are involved; activation of free myristate to myristoyl-CoA and transfer of the myristoyl residue to the Gag N-terminal glycine. We have investigated the effects of triacsin C, an inhibitor of long chain acyl-CoA synthetase, on release of HIV Gag virus-like particle (VLP) produced using the recombinant baculovirus system. First, inhibition of acyl-CoA formation by triacsin C was confirmed using the membrane fractions of insect Sf9 cells as an enzyme source. Second, when HIV Gag protein was expressed in the presence of triacsin C (0-48 microM), Gag myristoylation was inhibited in a dose-dependent manner. Budding of Gag VLP, however, did not follow similar inhibition kinetics but appeared unaffected up to 24 microM, yet was completely abolished at 48 microM when the myristoylation of Gag protein was also completely inhibited. The "all-or-none" inhibition of Gag VLP budding suggests that although inhibition of acyl-CoA synthetase blocks the production of myristoylated Gag protein, only complete inhibition of Gag myristoylation prevents VLP budding. Thus, relatively few myristoylated Gag molecules are sufficient for plasma membrane targeting and VLP budding.
Morikawa
Y
Y
Kitasato Institute, Tokyo, Japan.
Hinata
S
S
Tomoda
H
H
Goto
T
T
Nakai
M
M
Aizawa
C
C
Tanaka
H
H
Omura
S
S
D000214:Acyl Coenzyme A / Q000096:biosynthesis; D000818:Animals; D002460:Cell Line; D003066:Coenzyme A Ligases / Q000037:antagonists & inhibitors; D004791:Enzyme Inhibitors / Q000494:pharmacology; D015683:Gene Products, gag / Q000235:genetics / Q000378:metabolism*; D006678:HIV / Q000378:metabolism*; D008854:Microscopy, Electron; D019814:Myristic Acid; D009227:Myristic Acids / Q000037:antagonists & inhibitors*; D017924:Nucleopolyhedroviruses / Q000235:genetics; D012097:Repressor Proteins; D029701:Saccharomyces cerevisiae Proteins; D018411:Spodoptera; D014226:Triazenes / Q000494:pharmacology; D014771:Virion / Q000187:drug effects / Q000502:physiology; D000098859:Long-Chain-Fatty-Acid-CoA Ligase
D000214:Acyl Coenzyme A; D004791:Enzyme Inhibitors; D015683:Gene Products, gag; D009227:Myristic Acids; D012097:Repressor Proteins; D029701:Saccharomyces cerevisiae Proteins; D014226:Triazenes; D019814:Myristic Acid; C040349:S-tetradecanoyl-coenzyme A; C034613:triacsin C; D003066:Coenzyme A Ligases; C090799:FAA2 protein, S cerevisiae; D000098859:Long-Chain-Fatty-Acid-CoA Ligase