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Journal of Virology, December 2002, p. 11845-11852, Vol. 76, No. 23
0022-538X/02/$04.00+0     DOI: 10.1128/JVI.76.23.11845-11852.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Palmitoylation of the Murine Leukemia Virus Envelope Protein Is Critical for Lipid Raft Association and Surface Expression

Min Li, Chinglai Yang, Suxiang Tong,{dagger} Armin Weidmann,{ddagger} and Richard W. Compans*

Department of Microbiology and Immunology and Emory Vaccine Center, Emory University School of Medicine, Atlanta, Georgia 30322

Received 28 May 2002/ Accepted 23 August 2002

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-ß-cyclodextrin before extraction with ice-cold TX-100 or when treated with ice-cold octyl-ß-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.

* Corresponding author. Mailing address: Department of Microbiology and Immunology, Emory University School of Medicine, 3001 Rollins Research Center, Atlanta, GA 30322. Phone: (404) 727-5947. Fax: (404) 727-8250. E-mail: compans{at}microbio.emory.edu.

{dagger} Present address: Centers for Disease Control and Prevention, Atlanta, GA 30333.

{ddagger} Present address: MorphoSys AG, Research & Development, 82152 Martinsried/Planegg, Germany.

Journal of Virology, December 2002, p. 11845-11852, Vol. 76, No. 23
0022-538X/02/$04.00+0     DOI: 10.1128/JVI.76.23.11845-11852.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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