Characterization of functional hepatitis C virus envelope glycoproteins.
Hepatitis C virus (HCV) encodes two envelope glycoproteins, E1 and E2, that assemble as a noncovalent heterodimer which is mainly retained in the endoplasmic reticulum. Because assembly into particles and secretion from the cell lead to structural changes in viral envelope proteins, characterization of the proteins associated with the virion is necessary in order to better understand how they mature to be functional in virus entry. There is currently no efficient and reliable cell culture system to amplify HCV, and the envelope glycoproteins associated with the virion have therefore not been characterized yet. Recently, infectious pseudotype particles that are assembled by displaying unmodified HCV envelope glycoproteins on retroviral core particles have been successfully generated. Because HCV pseudotype particles contain fully functional envelope glycoproteins, these envelope proteins, or at least a fraction of them, should be in a mature conformation similar to that on the native HCV particles. In this study, we used conformation-dependent monoclonal antibodies to characterize the envelope glycoproteins associated with HCV pseudotype particles. We showed that the functional unit is a noncovalent E1E2 heterodimer containing complex or hybrid type glycans. We did not observe any evidence of maturation by a cellular endoprotease during the transport of these envelope glycoproteins through the secretory pathway. These envelope glycoproteins were recognized by a panel of conformation-dependent monoclonal antibodies as well as by CD81, a molecule involved in HCV entry. The functional envelope glycoproteins associated with HCV pseudotype particles were also shown to be sensitive to low-pH treatment. Such conformational changes are likely necessary to initiate fusion.
CNRS-UPR2511, Institut de Biologie de Lille and Institut Pasteur de Lille, Lille, France.
Anne
A
Op De Beeck
Cécile
C
Voisset
Birke
B
Bartosch
Yann
Y
Ciczora
Laurence
L
Cocquerel
Zhenyong
Z
Keck
Steven
S
Foung
François-Loïc
FL
Cosset
Jean
J
Dubuisson
D000911:Antibodies, Monoclonal; D002460:Cell Line; D019281:Dimerization; D016174:Hepacivirus / Q000378:metabolism*; D006801:Humans; D011487:Protein Conformation; D014759:Viral Envelope Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D015678:Viral Structural Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D014771:Virion / Q000378:metabolism*
D000911:Antibodies, Monoclonal; D014759:Viral Envelope Proteins; D015678:Viral Structural Proteins; C107842:protein E1, Classical swine fever virus; C091015:glycoprotein E2, Hepatitis C virus