title: Arenavirus entry occurs through a cholesterol-dependent, non-caveolar, clathrin-mediated endocytic mechanism.
abstract: Arenaviruses are important causes of viral hemorrhagic fevers in humans. Arenavirus infection of cells occurs via a pH-dependent endocytic route, but detailed studies of entry pathways have not been done. We investigated the role of cell membrane cholesterol, caveolae, and clathrin coated pits in infection by Lassa virus (LASV), which utilizes alpha-dystroglycan (alpha-DG) as a receptor, and Pichindé virus (PICV), which does not. Depletion of cellular cholesterol by treatment with methyl betacyclodextrin (MbetaCD) or nystatin/progesterone inhibited PICV replication and transfer of packaged marker gene by LASV or PICV pseudotyped retroviral particles. In cells lacking caveolae due to silencing of the caveolin-1 gene, no inhibition of PICV infection or LASV pseudotype transduction was observed. However, PICV infection and LASV and PICV pseudotype transduction was inhibited when an Eps15 dominant negative mutant was used to inhibit clathrin-mediated endocytosis. Altogether, the results indicate that diverse arenaviruses have a common requirement for cell membrane cholesterol and clathrin mediated endocytosis in establishing infection.
authors: Department of Pathology, Center for Biodefense and Emerging Infectious Diseases, The University of Texas Medical Branch, 301 University Blvd, Galveston, TX 77555, USA. emvela@utmb.edu
authors: Eric M
authors: EM
authors: Vela
authors: Lihong
authors: L
authors: Zhang
authors: Tonya M
authors: TM
authors: Colpitts
authors: Robert A
authors: RA
authors: Davey
authors: Judith F
authors: JF
authors: Aronson
mesh_terms: D000818:Animals; D021941:Caveolae / Q000821:virology; D051242:Caveolin 1 / Q000037:antagonists & inhibitors; D002460:Cell Line; D002522:Chlorocebus aethiops; D002784:Cholesterol / Q000096:biosynthesis; D022163:Clathrin-Coated Vesicles / Q000821:virology*; D004705:Endocytosis / Q000502:physiology*; D020868:Gene Silencing; D006801:Humans; D007836:Lassa virus / Q000502:physiology*; D008563:Membrane Lipids / Q000502:physiology*; D051379:Mice; D018052:Pichinde virus / Q000502:physiology*; D014161:Transduction, Genetic; D014709:Vero Cells; D053586:Virus Internalization; D014779:Virus Replication / Q000502:physiology
chemical_list: D051242:Caveolin 1; D008563:Membrane Lipids; D002784:Cholesterol
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