title: HS-AFM and SERS Analysis of Murine Norovirus Infection: Involvement of the Lipid Rafts.
abstract: Studies on human norovirus are severely hampered by the absence of a cell culture system until the discovery of murine norovirus (MNV). The cell membrane domains called lipid rafts have been defined as a port of entry for viruses. This study is conducted to investigate murine norovirus binding on the mouse leukemic monocyte macrophage cell line. Lipid raft related structures are extracted from cells by detergent treatment resulting detergent-resistant membrane (DRMs) domains. The real-time polymerase chain reaction technique is performed to detect the viral genome, thereby the MNV binding on the DRMs. The interactions between MNV and DRMs are investigated by high-speed atomic force microscopy (HS-AFM) combined with surface-enhanced Raman spectroscopy (SERS). The inoculation of the virus onto cells results in the aggregations of detergent-resistant membrane domains significantly. The characteristic Raman band of MNV is found in inoculated samples. To be sure that these results are originated from specific interactions between DRM and MNV, methyl-β-cyclo-dextrin (MβCD) is applied to disrupt lipid rafts. The MNV binding on DRMs is precluded by the MβCD treatment. The cholesterols chains are defined as a key factor in the interactions between norovirus and DRMs. The authors conclude that the MNV binding involves the presence of DRMs and cholesterol dependent.
authors.affiliation: ICB UMR CNRS 6303, University of Bourgogne Franche-Comte, Dijon, F-21000, France.
authors.forename: Ece N
authors.initials: EN
authors.lastname: Aybeke
authors.affiliation: Laboratory of Virology, National Reference Center for Enteric Viruses, CHU F. Mitterrand, Dijon, F-21000, France.|AgroSup Dijon, PAM UMR A 02.102, University of Bourgogne Franche-Comte, Dijon, F-21000, France.
authors.forename: Gaël
authors.initials: G
authors.lastname: Belliot
authors.affiliation: Lipid Nutrition Cancer, INSERM UMR 866, CHU F. Mitterrand, Dijon, F-21000, France.
authors.forename: Stéphanie
authors.initials: S
authors.lastname: Lemaire-Ewing
authors.affiliation: Laboratory of Virology, National Reference Center for Enteric Viruses, CHU F. Mitterrand, Dijon, F-21000, France.|AgroSup Dijon, PAM UMR A 02.102, University of Bourgogne Franche-Comte, Dijon, F-21000, France.
authors.forename: Marie
authors.initials: M
authors.lastname: Estienney
authors.affiliation: ICB UMR CNRS 6303, University of Bourgogne Franche-Comte, Dijon, F-21000, France.
authors.forename: Yvon
authors.initials: Y
authors.lastname: Lacroute
authors.affiliation: Laboratory of Virology, National Reference Center for Enteric Viruses, CHU F. Mitterrand, Dijon, F-21000, France.|AgroSup Dijon, PAM UMR A 02.102, University of Bourgogne Franche-Comte, Dijon, F-21000, France.
authors.forename: Pierre
authors.initials: P
authors.lastname: Pothier
authors.affiliation: ICB UMR CNRS 6303, University of Bourgogne Franche-Comte, Dijon, F-21000, France.
authors.forename: Eric
authors.initials: E
authors.lastname: Bourillot
authors.affiliation: ICB UMR CNRS 6303, University of Bourgogne Franche-Comte, Dijon, F-21000, France.
authors.forename: Eric
authors.initials: E
authors.lastname: Lesniewska
mesh_terms: D000818:Animals; D017250:Caliciviridae Infections / Q000378:metabolism*; D021962:Membrane Microdomains / Q000187:drug effects / Q000378:metabolism*; D051379:Mice; D018625:Microscopy, Atomic Force / Q000379:methods*; D029322:Norovirus / Q000502:physiology*; D000067996:RAW 264.7 Cells; D060888:Real-Time Polymerase Chain Reaction; D013059:Spectrum Analysis, Raman / Q000379:methods*; D047392:beta-Cyclodextrins / Q000494:pharmacology
chemical_list: D047392:beta-Cyclodextrins; C031215:betadex
keywords: detergent-resistant membrane domains; high-speed atomic force microscopy; lipid rafts; norovirus; surface enhanced Raman spectroscopy