title: Identification of interactions in the E1E2 heterodimer of hepatitis C virus important for cell entry.
abstract: Several conserved domains critical for E1E2 assembly and hepatitis C virus entry have been identified in E1 and E2 envelope glycoproteins. However, the role of less conserved domains involved in cross-talk between either glycoprotein must be defined to fully understand how E1E2 undergoes conformational changes during cell entry. To characterize such domains and to identify their functional partners, we analyzed a set of intergenotypic E1E2 heterodimers derived from E1 and E2 of different genotypes. The infectivity of virions indicated that Con1 E1 did not form functional heterodimers when associated with E2 from H77. Biochemical analyses demonstrated that the reduced infectivity was not related to alteration of conformation and incorporation of Con1 E1/H77 E2 heterodimers but rather to cell entry defects. Thus, we generated chimeric E1E2 glycoproteins by exchanging different domains of each protein in order to restore functional heterodimers. We found that both the ectodomain and transmembrane domain of E1 influenced infectivity. Site-directed mutagenesis highlighted the role of amino acids 359, 373, and 375 in transmembrane domain in entry. In addition, we identified one domain involved in entry within the N-terminal part of E1, and we isolated a motif at position 219 that is critical for H77 function. Interestingly, using additional chimeric E1E2 complexes harboring substitutions in this motif, we found that the transmembrane domain of E1 acts as a partner of this motif. Therefore, we characterized domains of E1 and E2 that have co-evolved inside a given genotype to optimize their interactions and allow efficient entry.
authors.affiliation: Université de Lyon, UCB-Lyon1, IFR128, INSERM, U758, and École Normale Supérieure de Lyon, Lyon F-69007, France.
authors.forename: Guillemette
authors.initials: G
authors.lastname: Maurin
authors.forename: Judith
authors.initials: J
authors.lastname: Fresquet
authors.forename: Ophélia
authors.initials: O
authors.lastname: Granio
authors.forename: Czeslaw
authors.initials: C
authors.lastname: Wychowski
authors.forename: François-Loïc
authors.initials: FL
authors.lastname: Cosset
authors.forename: Dimitri
authors.initials: D
authors.lastname: Lavillette
mesh_terms: D020816:Amino Acid Motifs; D002460:Cell Line; D016174:Hepacivirus / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D020836:Protein Structure, Quaternary; D017434:Protein Structure, Tertiary; D014759:Viral Envelope Proteins / Q000235:genetics / Q000378:metabolism*; D053586:Virus Internalization
chemical_list: C087035:E1 protein, Hepatitis C virus; D014759:Viral Envelope Proteins; C091015:glycoprotein E2, Hepatitis C virus