Core protein domains involved in hepatitis C virus-like particle assembly and budding at the endoplasmic reticulum membrane.
Hepatitis C virus (HCV) core protein, expressed with a Semliki forest virus (SFV) replicon, self-assembles into HCV-like particles (HCV-LPs) at the endoplasmic reticulum (ER) membrane, providing an opportunity to study HCV particle morphogenesis by electron microscopy. Various mutated HCV core proteins with engineered internal deletions were expressed with this system, to identify core domains required or dispensable for HCV-LP assembly. The HCV core protein sequence was compared with its counterpart in GB virus B (GBV-B), the virus most closely related to HCV, to identify conserved domains. GBV-B and HCV display similar tropism for liver hepatocytes and their core proteins are organized similarly into three main domains (I, II and III), although GBV-B core is smaller and lacks approximately 35 amino acids (aa) in domain I. The deletion of short hydrophobic domains (aa 133-152 and 153-167 in HCV core) that appear highly conserved in domain II of both GBV-B and HCV core proteins resulted in loss of HCV core ER anchoring and self-assembly into HCV-LPs. The deletion of short domains found within domain I of HCV core protein but not in the corresponding domain of GBV-B core according to sequence alignment had contrasting effects. Amino acids 15-28 and 60-66 were shown to be dispensable for HCV-LP assembly and morphogenesis, whereas aa 88-106 were required for this process. The production of GBV-B core protein from a recombinant SFV vector was associated with specific ER ultrastructural changes, but did not lead to the morphogenesis of GBV-B-LPs, suggesting that different budding mechanisms occur in members of the Flaviviridae family.
Hourioux
Christophe
C
Université François Rabelais de Tours, INSERM ERI 19 Tours, France.
Ait-Goughoulte
Malika
M
Patient
Romuald
R
Fouquenet
Delphine
D
Arcanger-Doudet
Fabienne
F
Brand
Denys
D
Martin
Annette
A
Roingeard
Philippe
P
D000595:Amino Acid Sequence; D000818:Animals; D015153:Blotting, Western; D002460:Cell Line; D004721:Endoplasmic Reticulum / Q000378:metabolism* / Q000648:ultrastructure / Q000821:virology; D029406:GB virus B / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D016174:Hepacivirus / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D057927:Hydrophobic and Hydrophilic Interactions; D018613:Microscopy, Confocal; D046529:Microscopy, Electron, Transmission; D008969:Molecular Sequence Data; D009154:Mutation; D017384:Sequence Deletion; D017386:Sequence Homology, Amino Acid; D014758:Viral Core Proteins / Q000737:chemistry / Q000235:genetics / Q000502:physiology*
D014758:Viral Core Proteins