Identification of a conserved residue of foamy virus Gag required for intracellular capsid assembly.
In contrast to all retroviruses but similar to the hepatitis B virus, foamy viruses (FV) require expression of the envelope protein for budding of intracellular capsids from the cell, suggesting a specific interaction between the Gag and Env proteins. Capsid assembly occurs in the cytoplasm of infected cells in a manner similar to that for the B- and D-type viruses; however, in contrast to these retroviruses, FV Gag lacks an N-terminal myristylation signal and capsids are not targeted to the plasma membrane (PM). We have found that mutation of an absolutely conserved arginine (Arg) residue at position 50 to alanine (R50A) of the simian foamy virus SFV cpz(hu) inhibits proper capsid assembly and abolishes viral budding even in the presence of the envelope (Env) glycoproteins. Particle assembly and extracellular release of virus can be restored to this mutant with the addition of an N-terminal Src myristylation signal (Myr-R50A), presumably by providing an alternate site for assembly to occur at the PM. In addition, the strict requirement of Env expression for capsid budding can be bypassed by addition of a PM-targeting signal to Gag. These results suggest that intracellular capsid assembly may be mediated by a signal akin to the cytoplasmic targeting and retention signal CTRS found in Mason-Pfizer monkey virus and that FV Gag has the inherent ability to assemble capsids at multiple sites like conventional retroviruses. The necessity of Env expression for particle egress is most probably due to the lack of a membrane-targeting signal within FV Gag to direct capsids to the PM for release and indicates that Gag-Env interactions are essential to drive particle budding.
Eastman
S W
SW
Division of Basic Sciences, Fred Hutchinson Cancer Research Center, Seattle, Washington 98109, USA.
Linial
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ML
D000595:Amino Acid Sequence; D001120:Arginine / Q000235:genetics; D002213:Capsid / Q000235:genetics / Q000502:physiology*; D002462:Cell Membrane / Q000378:metabolism; D017124:Conserved Sequence; D015683:Gene Products, gag / Q000235:genetics / Q000502:physiology*; D006801:Humans; D007424:Intracellular Fluid; D008969:Molecular Sequence Data; D017354:Point Mutation; D016092:Spumavirus / Q000235:genetics / Q000502:physiology*; D013347:Subcellular Fractions; D014771:Virion; D019065:Virus Assembly
D015683:Gene Products, gag; D001120:Arginine