Effects of disulfide bridges glycoprotein E1 on fusogenic activity of Rubella virus.
Rubella virus (RUBV) infects cells via an acid-triggered membrane fusion process. RUBV virions contain two cysteine-rich glycoproteins, E2 and E1. The latter is believed to be involved in the membrane fusion. Using a recombinant plasmid containing RUBV E1 and E2, 11 of total 20 cysteines present in the ectodomain of wild type E1 were mutated to test their role in the fusion via the formation of disulfide bridges. The recombinant plasmids containing mutated E1 (Cys2-Cys20) or wild type (wt) E1 were expressed in BHK-21 cells. Their fusogenic and hemadsorption activities in addition to a potential of cell surface expression of E1 and E2 were assayed. The results showed that the fusogenic activity was lost in all tested mutants, while the hemadsorption activity and cell surface expression potential were affected differently in individual mutants. Since only the Cys5 and Cys8 mutations led to a reduction of both hemadsorption and cell surface expression, we assume that these mutations prevented the formation of the disulfide bridge, what led to a misfolding of E1 and consequently to a failure of recognition of E1 by E2. In conclusion, the disulfide bridges disrupted in all the tested mutants appear essential for the cell fusion, while only the disulfide bridge C(5)-C(8) seems to be crucial for the transport of E1 and E2 in the cell.
Liu
X L
XL
Department of Virology, Shandong University, Jinan, P.R. China.
Wu
B
B
Zhang
W Q
WQ
Song
Y Y
YY
Xu
H Z
HZ
Wang
G T
GT
Wang
Z Y
ZY
D019943:Amino Acid Substitution; D000818:Animals; D002460:Cell Line; D006224:Cricetinae; D003545:Cysteine / Q000235:genetics; D004220:Disulfides / Q000737:chemistry*; D006382:Hemadsorption; D016297:Mutagenesis, Site-Directed; D021381:Protein Transport; D012412:Rubella virus / Q000737:chemistry* / Q000502:physiology*; D014759:Viral Envelope Proteins / Q000737:chemistry* / Q000235:genetics / Q000502:physiology*; D053586:Virus Internalization
D004220:Disulfides; D014759:Viral Envelope Proteins; C061826:E1 envelope protein, Rubella virus; D003545:Cysteine