title: Apolipoprotein E likely contributes to a maturation step of infectious hepatitis C virus particles and interacts with viral envelope glycoproteins.
abstract: The assembly of infectious hepatitis C virus (HCV) particles is tightly linked to components of the very-low-density lipoprotein (VLDL) pathway. We and others have shown that apolipoprotein E (ApoE) plays a major role in production of infectious HCV particles. However, the mechanism by which ApoE contributes to virion assembly/release and how it gets associated with the HCV particle is poorly understood. We found that knockdown of ApoE reduces titers of infectious intra- and extracellular HCV but not of the related dengue virus. ApoE depletion also reduced amounts of extracellular HCV core protein without affecting intracellular core amounts. Moreover, we found that ApoE depletion affected neither formation of nucleocapsids nor their envelopment, suggesting that ApoE acts at a late step of assembly, such as particle maturation and infectivity. Importantly, we demonstrate that ApoE interacts with the HCV envelope glycoproteins, most notably E2. This interaction did not require any other viral proteins and depended on the transmembrane domain of E2 that also was required for recruitment of HCV envelope glycoproteins to detergent-resistant membrane fractions. These results suggest that ApoE plays an important role in HCV particle maturation, presumably by direct interaction with viral envelope glycoproteins.
  
  
  OBJECTIVE
  The HCV replication cycle is tightly linked to host cell lipid pathways and components. This is best illustrated by the dependency of HCV assembly on lipid droplets and the VLDL component ApoE. Although the role of ApoE for production of infectious HCV particles is well established, it is still poorly understood how ApoE contributes to virion formation and how it gets associated with HCV particles. Here, we provide experimental evidence that ApoE likely is required for an intracellular maturation step of HCV particles. Moreover, we demonstrate that ApoE associates with the viral envelope glycoproteins. This interaction appears to be dispensable for envelopment of virus particles but likely contributes to the quality control of secreted infectious virions. These results shed new light on the exploitation of host cell lipid pathways by HCV and the link of viral particle assembly to the VLDL component ApoE.
authors.affiliation: Department of Infectious Diseases, Molecular Virology, University Hospital Heidelberg, Heidelberg, Germany.
authors.forename: Ji-Young
authors.initials: JY
authors.lastname: Lee
authors.affiliation: Department of Infectious Diseases, Molecular Virology, University Hospital Heidelberg, Heidelberg, Germany.
authors.forename: Eliana G
authors.initials: EG
authors.lastname: Acosta
authors.affiliation: Department of Infectious Diseases, Molecular Virology, University Hospital Heidelberg, Heidelberg, Germany.
authors.forename: Ina Karen
authors.initials: IK
authors.lastname: Stoeck
authors.affiliation: Department of Infectious Diseases, Molecular Virology, University Hospital Heidelberg, Heidelberg, Germany.
authors.forename: Gang
authors.initials: G
authors.lastname: Long
authors.affiliation: Department of Infectious Diseases, Molecular Virology, University Hospital Heidelberg, Heidelberg, Germany.
authors.forename: Marie-Sophie
authors.initials: MS
authors.lastname: Hiet
authors.affiliation: Department of Infectious Diseases, Integrative Virology, University Hospital Heidelberg, Heidelberg, Germany.
authors.forename: Birthe
authors.initials: B
authors.lastname: Mueller
authors.affiliation: Department of Infectious Diseases, Integrative Virology, University Hospital Heidelberg, Heidelberg, Germany.
authors.forename: Oliver T
authors.initials: OT
authors.lastname: Fackler
authors.affiliation: Department of Infectious Diseases, Molecular Virology, University Hospital Heidelberg, Heidelberg, Germany.
authors.forename: Stephanie
authors.initials: S
authors.lastname: Kallis
authors.affiliation: Department of Infectious Diseases, Molecular Virology, University Hospital Heidelberg, Heidelberg, Germany Ralf_Bartenschlager@med.uni-heidelberg.de.
authors.forename: Ralf
authors.initials: R
authors.lastname: Bartenschlager
mesh_terms: D001057:Apolipoproteins E / Q000378:metabolism*; D002460:Cell Line; D016174:Hepacivirus / Q000502:physiology*; D054884:Host-Pathogen Interactions; D006801:Humans; D011485:Protein Binding; D014759:Viral Envelope Proteins / Q000378:metabolism*; D019065:Virus Assembly
chemical_list: D001057:Apolipoproteins E; D014759:Viral Envelope Proteins; C091015:glycoprotein E2, Hepatitis C virus