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      Quantitative analysis of the lipidomes of the influenza virus envelope and MDCK cell apical membrane

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          Abstract

          Analysis of the lipid composition of influenza virus–infected cells provides support for the membrane raft-based biogenesis model.

          Abstract

          The influenza virus (IFV) acquires its envelope by budding from host cell plasma membranes. Using quantitative shotgun mass spectrometry, we determined the lipidomes of the host Madin–Darby canine kidney cell, its apical membrane, and the IFV budding from it. We found the apical membrane to be enriched in sphingolipids (SPs) and cholesterol, whereas glycerophospholipids were reduced, and storage lipids were depleted compared with the whole-cell membranes. The virus membrane exhibited a further enrichment of SPs and cholesterol compared with the donor membrane at the expense of phosphatidylcholines. Our data are consistent with and extend existing models of membrane raft-based biogenesis of the apical membrane and IFV envelope.

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          Most cited references46

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          Virus entry by endocytosis.

          Although viruses are simple in structure and composition, their interactions with host cells are complex. Merely to gain entry, animal viruses make use of a repertoire of cellular processes that involve hundreds of cellular proteins. Although some viruses have the capacity to penetrate into the cytosol directly through the plasma membrane, most depend on endocytic uptake, vesicular transport through the cytoplasm, and delivery to endosomes and other intracellular organelles. The internalization may involve clathrin-mediated endocytosis (CME), macropinocytosis, caveolar/lipid raft-mediated endocytosis, or a variety of other still poorly characterized mechanisms. This review focuses on the cell biology of virus entry and the different strategies and endocytic mechanisms used by animal viruses.
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            The HIV lipidome: a raft with an unusual composition.

            The lipids of enveloped viruses play critical roles in viral morphogenesis and infectivity. They are derived from the host membranes from which virus budding occurs, but the precise lipid composition has not been determined for any virus. Employing mass spectrometry, this study provides a quantitative analysis of the lipid constituents of HIV and a comprehensive comparison with its host membranes. Both a substantial enrichment of the unusual sphingolipid dihydrosphingomyelin and a loss of viral infectivity upon inhibition of sphingolipid biosynthesis in host cells are reported, establishing a critical role for this lipid class in the HIV replication cycle. Intriguingly, the overall lipid composition of native HIV membranes resembles detergent-resistant membrane microdomains and is strikingly different from that of host cell membranes. With this composition, the HIV lipidome provides strong evidence for the existence of lipid rafts in living cells.
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              A novel informatics concept for high-throughput shotgun lipidomics based on the molecular fragmentation query language

              Shotgun lipidome profiling relies on direct mass spectrometric analysis of total lipid extracts from cells, tissues or organisms and is a powerful tool to elucidate the molecular composition of lipidomes. We present a novel informatics concept of the molecular fragmentation query language implemented within the LipidXplorer open source software kit that supports accurate quantification of individual species of any ionizable lipid class in shotgun spectra acquired on any mass spectrometry platform.

                Author and article information

                Journal
                J Cell Biol
                J. Cell Biol
                jcb
                The Journal of Cell Biology
                The Rockefeller University Press
                0021-9525
                1540-8140
                23 January 2012
                : 196
                : 2
                : 213-221
                Affiliations
                [1 ]Max Planck Institute of Molecular Cell Biology and Genetics, 01307 Dresden, Germany
                [2 ]Research Institute, Division of Molecular Structure and Function, The Hospital for Sick Children, Toronto, Ontario M5G 1X8, Canada
                [3 ]Institut für Virologie, Medizinische Fakultät Carl Gustav Carus, Technische Universität Dresden, 01062 Dresden, Germany
                [4 ]Department of Biochemistry and [5 ]Department of Laboratory Medicine and Pathobiology, University of Toronto, Toronto, Ontario M5S 1A1, Canada
                Author notes
                Correspondence to Kai Simons: simons@ 123456mpi-cbg.de

                M.J. Gerl and J.L. Sampaio contributed equally to this paper.

                M.J. Gerl’s and S. Urban’s present address is Heidelberg University Biochemistry Center, 69120 Heidelberg, Germany.

                L. Kalvodova’s present address is Life Sciences, Wiley-VCH, 69469 Weinheim, Germany.

                Article
                201108175
                10.1083/jcb.201108175
                3265945
                22249292
                85e82a86-0f3a-4eb5-bd6a-0cb83a55ae50
                © 2012 Gerl et al.

                This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 3.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/3.0/).

                History
                : 30 August 2011
                : 16 December 2011
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                Cell biology
                Cell biology

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