title: Hepatitis E virus enters liver cells through a dynamin-2, clathrin and membrane cholesterol-dependent pathway.
abstract: The hepatitis E virus (HEV) causes large outbreaks and sporadic cases of acute viral hepatitis in developing countries. In the developed world, HEV occurrence has increased as a result of zoonotic transmission from swine. The cellular aspects of HEV infection, especially the determinants of entry, are poorly understood. In the absence of a robust in vitro culture system for HEV, it is not possible to produce high titre infectious virus that can be labeled for tracking its internalization. We have therefore used an Escherichia coli expressed HEV-like particle (HEV-LP) to study HEV entry. Following internalization, the HEV-LP initially trafficks to Rab5-positive compartments en route to acidic lysosomal compartments where it is degraded. Using pharmacological inhibitors, dominant negative and constitutively active mutants, and siRNA-mediated perturbations, we show that HEV entry requires dynamin-2, clathrin, membrane cholesterol and actin, but is independent of factors associated with macropinocytosis. The HEV-LP results were further validated through infection of liver cells with virus from the stool of an infected patient. The comparative analysis also showed involvement of the phosphatidylinositol-3-kinase/Akt pathway in an early post-entry step of viral replication. This report provides a detailed description of endocytic processes associated with HEV infection.
authors.affiliation: Virology Group, International Centre for Genetic Engineering Biotechnology, New Delhi, India.
authors.forename: Prasida
authors.initials: P
authors.lastname: Holla
authors.forename: Imran
authors.initials: I
authors.lastname: Ahmad
authors.forename: Zulfazal
authors.initials: Z
authors.lastname: Ahmed
authors.forename: Shahid
authors.initials: S
authors.lastname: Jameel
mesh_terms: D000199:Actins / Q000378:metabolism; D002460:Cell Line; D045744:Cell Line, Tumor; D002784:Cholesterol / Q000378:metabolism*; D002966:Clathrin / Q000378:metabolism*; D034285:Dynamin II / Q000378:metabolism*; D004926:Escherichia coli / Q000378:metabolism; D057809:HEK293 Cells; D006367:HeLa Cells; D016752:Hepatitis E virus / Q000502:physiology*; D022781:Hepatocytes / Q000378:metabolism / Q000821:virology*; D006801:Humans; D008099:Liver / Q000378:metabolism / Q000821:virology*; D058539:Phosphatidylinositol 3-Kinase / Q000378:metabolism; D010873:Pinocytosis / Q000502:physiology; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism; D015398:Signal Transduction / Q000502:physiology; D053586:Virus Internalization; D014779:Virus Replication / Q000502:physiology
chemical_list: D000199:Actins; D002966:Clathrin; D002784:Cholesterol; D058539:Phosphatidylinositol 3-Kinase; D051057:Proto-Oncogene Proteins c-akt; D034285:Dynamin II
keywords: PI3 kinase; actin; cholesterol; clathrin; dynamin; hepatitis; hepatitis E virus; virus-like particle