The transmembrane domain of the hepatitis C virus E2 glycoprotein is required for correct folding of the E1 glycoprotein and native complex formation.
Hepatitis C virus (HCV) encodes two glycoproteins, E1 and E2, that interact to form both native and aggregated complexes in tissue culture cells. In native complexes, E1 and E2 are associated by noncovalent interactions and such complexes are considered to constitute the authentic interactions between the proteins. By contrast, the proteins are linked by covalent, disulfide bonds in aggregated complexes. From studies with a mutant in which cysteine residues in E1 have been substituted with other amino acids, we show that E1 continues to associate with E2, although the migratory patterns of the proteins on gels are consistent with the formation of aggregated complexes. Therefore, such complexes can be stabilized by noncovalent as well as covalent interactions. To further examine the requirements for native complex formation, segments of foreign glycoproteins were linked to regions of E2. Our data provide direct evidence for the requirement of C-terminal sequences in E2 that contain the transmembrane domain to permit oxidation of E1 and assembly of a native complex. By contrast, native complexes and oxidized E1 are not found in the presence of chimeric proteins containing the E2 ectodomain. These data suggest that interaction of E1 with the E2 transmembrane domain is critical for native complex formation.
Patel
J
J
MRC Virology Unit, Institute of Virology, Church Street, Glasgow, G11 5JR, United Kingdom.
Patel
A H
AH
McLauchlan
J
J
D000818:Animals; D015153:Blotting, Western; D002460:Cell Line; D002462:Cell Membrane / Q000737:chemistry; D006224:Cricetinae; D004591:Electrophoresis, Polyacrylamide Gel; D018274:Electroporation; D019084:Fluorescent Antibody Technique, Indirect; D010084:Oxidation-Reduction; D011233:Precipitin Tests; D011487:Protein Conformation; D017510:Protein Folding; D017434:Protein Structure, Tertiary; D014158:Transcription, Genetic; D014759:Viral Envelope Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*
C087035:E1 protein, Hepatitis C virus; D014759:Viral Envelope Proteins; C091015:glycoprotein E2, Hepatitis C virus