Palmitoylation of the murine leukemia virus envelope protein is critical for lipid raft association and surface expression.
To investigate the association of the murine leukemia virus (MuLV) Env protein with lipid rafts, we compared wild-type and palmitoylation-deficient mutant Env proteins by using extraction with the mild detergent Triton X-100 (TX-100) followed by a sucrose gradient flotation assay. We found that the wild-type MuLV Env protein was resistant to ice-cold TX-100 treatment and floated to the top of the gradients. In contrast, we observed that the palmitoylation-deficient mutant Env protein was mostly soluble when extracted by ice-cold TX-100 and stayed at the bottom of the gradients. Both the wild-type and mutant Env proteins were found to be soluble when treated with methyl-beta-cyclodextrin before extraction with ice-cold TX-100 or when treated with ice-cold octyl-beta-glucoside instead of TX-100. These results indicate that the MuLV Env protein is associated with lipid rafts and that palmitoylation of the Env protein is critical for lipid raft association. Although the palmitoylation-deficient Env mutant was synthesized at a level similar to that of the wild-type Env, it was found to be expressed at reduced levels on the cell surface. We observed syncytium formation activity with both the wild-type and mutant Env proteins, indicating that palmitoylation or raft association is not required for MuLV viral fusion activity.
Li
Min
M
Department of Microbiology and Immunology and Emory Vaccine Center, Emory University School of Medicine, Atlanta, Georgia 30322, USA.
Yang
Chinglai
C
Tong
Suxiang
S
Weidmann
Armin
A
Compans
Richard W
RW
D016475:3T3 Cells; D000818:Animals; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism / Q000821:virology; D015726:Giant Cells / Q000473:pathology / Q000821:virology; D009052:Leukemia Virus, Murine / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D008561:Membrane Fusion; D021962:Membrane Microdomains / Q000378:metabolism*; D051379:Mice; D009154:Mutation; D017830:Octoxynol; D019308:Palmitic Acid / Q000378:metabolism*; D012995:Solubility; D014759:Viral Envelope Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*
D014759:Viral Envelope Proteins; D019308:Palmitic Acid; D017830:Octoxynol