Influenza Viruses Select Ordered Lipid Domains during Budding from the Plasma Membrane*
- From the Cell Biology Programme ‡Structural Biology Programme, European Molecular Biology Laboratory, Postfach 10 2209, D-69012 Heidelberg, Germany and Max-Planck-Institute for Molecular Cell Biology and Genetics, Dresden 01307, Germany
Abstract
During the budding of enveloped viruses from the plasma membrane, the lipids are not randomly incorporated into the envelope, but virions seem to have a lipid composition different from the host membrane. Here, we have analyzed lipid assemblies in three different viruses: fowl plague virus (FPV) from the influenza virus family, vesicular stomatitis virus (VSV), and Semliki Forest virus (SFV). Analysis of detergent extractability of proteins, cholesterol, phosphoglycerolipids, and sphingomyelin in virions showed that FPV contains high amounts of detergent-insoluble complexes, whereas such complexes are largely absent from VSV or SFV. Cholesterol depletion from the viral envelope by methyl-β-cyclodextrin results in increased solubility of sphingomyelin and of the glycoproteins in the FPV envelope. This biochemical behavior suggests that so-called raft-lipid domains are selectively incorporated into the influenza virus envelope. The “fluidity” of the FPV envelope, as measured by the fluorescence polarization of diphenylhexatriene, was significantly lower than compared with VSV or SFV. Furthermore, influenza virus hemagglutinin incorporated into the envelope of recombinant VSV was largely detergent-soluble, indicating the depletion of raft-lipid assemblies from this membrane. The results provide a model for lipid selectivity during virus budding and support the view of lipid rafts as cholesterol-dependent, ordered domains in biological membranes.
Footnotes
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↵* This work was supported by Deutsche Forschungsgemeinschaft Grant SFB 352 and the European Commission TMR program.The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked “advertisement” in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
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↵§ To whom correspondence should be addressed: European Molecular Biology Laboratory, Postfach 10 2209, D-69012 Heidelberg, Germany. Tel.: 6221-387-334; Fax: 6221-387-512; E-mail:Simons{at}emblHeidelberg.de.
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↵2 P. Scheiffele and K. Simons, unpublished data.
- Abbreviations:
- GPI
-
glycosylphosphatidylinositol
- DIG
-
detergent-insoluble glycolipid-enriched complex
- HA
-
hemagglutinin
- FPV
-
fowl plague virus
- VSV
-
vesicular stomatitis virus
- SFV
-
Semliki Forest virus
- BHK
-
bovine hamster kidney
- SM
-
sphingomyelin
- PC
-
phosphatidylcholine
- PE
-
phosphatidylethanolamine
- PAGE
-
polyacrylamide gel electrophoresis
- CD
-
methyl-β-cyclodextrin
- DPH
-
diphenylhexatriene
- DPPC
-
dipalmitoylphosphatidylcholine.
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- Received September 11, 1998.
- Revision received October 30, 1998.
- The American Society for Biochemistry and Molecular Biology, Inc.
