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      Cholesterol Dependence of Pseudorabies Herpesvirus Entry

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          Abstract

          Lipid rafts are special microdomains in the plasma membrane. They are enriched in sphingolipids and cholesterol, playing critical roles in many biological processes. The purpose of this study is to analyze the requirement of cholesterol, a crucial component of lipid rafts for cell infection by pseudorabies virus (PrV). Cholesterol of plasma membrane or viral envelope was depleted with methyl-beta-cyclodextrin (MβCD), and the infectivity of three strains of PrV was determined with plaque assays. The effect of adding cholesterol to MβCD-treated cells and viruses on cell infection was analyzed. Furthermore, effect of post-adsorption cholesterol depletion on PrV infection was investigated. We show that cholesterol depletion of either the plasma membrane or the viral membrane by MβCD significantly impaired the infectivity of PrV strains Kaplan, Becker, and Bartha K-61. The virus was shown to have lower cholesterol content and to respond to lower MβCD concentrations. Exogenous cholesterol added to either MβCD-treated cells or virions partially restored the virus infectivity. Optimal PrV infection requires cholesterol in viral and plasma membranes.

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          Lipid rafts and signal transduction.

          Signal transduction is initiated by complex protein-protein interactions between ligands, receptors and kinases, to name only a few. It is now becoming clear that lipid micro-environments on the cell surface -- known as lipid rafts -- also take part in this process. Lipid rafts containing a given set of proteins can change their size and composition in response to intra- or extracellular stimuli. This favours specific protein-protein interactions, resulting in the activation of signalling cascades.
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            Structure and function of sphingolipid- and cholesterol-rich membrane rafts.

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              Influenza viruses select ordered lipid domains during budding from the plasma membrane.

              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-beta-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.
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                Author and article information

                Contributors
                +86-451-55190385 , +86-451-55103336 , rxfemail@yahoo.com.cn , renxf@neau.edu.cn
                Journal
                Curr Microbiol
                Curr. Microbiol
                Current Microbiology
                Springer-Verlag (New York )
                0343-8651
                1432-0991
                13 July 2010
                2011
                : 62
                : 1
                : 261-266
                Affiliations
                [1 ]GRID grid.412243.2, ISNI 0000000417601136, College of Veterinary Medicine, , Northeast Agricultural University, ; 59 Mucai Street, Harbin, 150030 China
                [2 ]GRID grid.412243.2, ISNI 0000000417601136, College of Life Sciences, , Northeast Agricultural University, ; 59 Mucai Street, Harbin, 150030 China
                [3 ]GRID grid.412970.9, ISNI 0000000101266191, Institute for Virology, , University of Veterinary Medicine Hannover, ; Buenteweg 17, 30559 Hannover, Germany
                Article
                9700
                10.1007/s00284-010-9700-8
                7080178
                20625735
                © Springer Science+Business Media, LLC 2010

                This article is made available via the PMC Open Access Subset for unrestricted research re-use and secondary analysis in any form or by any means with acknowledgement of the original source. These permissions are granted for the duration of the World Health Organization (WHO) declaration of COVID-19 as a global pandemic.

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                © Springer Science+Business Media, LLC 2011

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