Ecotropic murine leukemia virus receptor is physically associated with caveolin and membrane rafts.
We used a Sindbis virus expression system to stably express a chimeric ecotropic murine leukemia virus (MLV) receptor gene, CAT1, fused to green fluorescent protein (gfp) in BHK cells. The chimeric gene was expressed on the cell surface and functioned as an MLV receptor. Using gfp as an epitope tag, we found that CAT1 cross-immunoprecipitated with caveolin, a cellular protein associated non-clathrin-coated endocytic vesicles. Biochemical studies showed that CAT1 copurified with caveolin in a detergent-insoluble membrane fraction that forms cholesterol-rich "rafts" on the cell surface. Disruption of rafts by methyl-beta-cyclodextrin, a drug that extracts cholesterol, reduced susceptibility to MLV without decreasing surface CAT1. The results indicate that association of the MLV receptor with cholesterol-rich rafts is important for an early step in virus infection.
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Laboratory of Molecular Microbiology, National Institutes of Health, Bethesda, Maryland, 20892, USA.
Silver
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J
D000818:Animals; D051242:Caveolin 1; D022461:Caveolins / Q000378:metabolism*; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism*; D002784:Cholesterol / Q000378:metabolism; D006224:Cricetinae; D008562:Membrane Glycoproteins / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D011991:Receptors, Virus / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D012845:Sindbis Virus
C495541:Cav1 protein, mouse; D051242:Caveolin 1; D022461:Caveolins; D008562:Membrane Glycoproteins; D011991:Receptors, Virus; D011993:Recombinant Fusion Proteins; C070506:ecotropic murine leukemia virus receptor; D002784:Cholesterol