title: Cholesterol-rich microdomains as docking platforms for respiratory syncytial virus in normal human bronchial epithelial cells.
abstract: Respiratory syncytial virus (RSV) is one of the major causes of respiratory infections in children, and it is the main pathogen causing bronchiolitis in infants. The binding and entry mechanism by which RSV infects respiratory epithelial cells has not yet been determined. In this study, the earliest stages of RSV infection in normal human bronchial epithelial cells were probed by tracking virions with fluorescent lipophilic dyes in their membranes. Virions colocalized with cholesterol-containing plasma membrane microdomains, identified by their ability to bind cholera toxin subunit B. Consistent with an important role for cholesterol in RSV infection, cholesterol depletion profoundly inhibited RSV infection, while cholesterol repletion reversed this inhibition. Merger of the outer leaflets of the viral envelope and the cell membrane appeared to be triggered at these sites. Using small-molecule inhibitors, RSV infection was found to be sensitive to Pak1 inhibition, suggesting the requirement of a subsequent step of cytoskeletal reorganization that could involve plasma membrane rearrangements or endocytosis. It appears that RSV entry depends on its ability to dock to cholesterol-rich microdomains (lipid rafts) in the plasma membrane where hemifusion events begin, assisted by a Pak1-dependent process.
authors.affiliation: Departamento de Medicina, Fundación Universidad del Norte, Barranquilla, Colombia. hsanjuan@uninorte.edu.co
authors.forename: Homero
authors.initials: H
authors.lastname: San-Juan-Vergara
authors.forename: Viviana
authors.initials: V
authors.lastname: Sampayo-Escobar
authors.forename: Niradiz
authors.initials: N
authors.lastname: Reyes
authors.forename: Byeong
authors.initials: B
authors.lastname: Cha
authors.forename: Lisandro
authors.initials: L
authors.lastname: Pacheco-Lugo
authors.forename: Terianne
authors.initials: T
authors.lastname: Wong
authors.forename: Mark E
authors.initials: ME
authors.lastname: Peeples
authors.forename: Peter L
authors.initials: PL
authors.lastname: Collins
authors.forename: Maria Eugenia
authors.initials: ME
authors.lastname: Castaño
authors.forename: Shyam S
authors.initials: SS
authors.lastname: Mohapatra
mesh_terms: D001980:Bronchi / Q000166:cytology / Q000378:metabolism*; D002460:Cell Line; D002784:Cholesterol / Q000378:metabolism*; D004847:Epithelial Cells / Q000378:metabolism; D005434:Flow Cytometry; D005456:Fluorescent Dyes; D006801:Humans; D008561:Membrane Fusion; D012136:Respiratory Syncytial Viruses / Q000502:physiology*
chemical_list: D005456:Fluorescent Dyes; D002784:Cholesterol
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