Poxvirus complement control proteins are expressed on the cell surface through an intermolecular disulfide bridge with the viral A56 protein.
The vaccinia virus (VACV) complement control protein (VCP) is an immunomodulatory protein that is both secreted from and expressed on the surface of infected cells. Surface expression of VCP occurs though an interaction with the viral transmembrane protein A56 and is dependent on a free N-terminal cysteine of VCP. Although A56 and VCP have been shown to interact in infected cells, the mechanism remains unclear. To investigate if A56 is sufficient for surface expression, we transiently expressed VCP and A56 in eukaryotic cell lines and found that they interact on the cell surface in the absence of other viral proteins. Since A56 contains three extracellular cysteines, we hypothesized that one of the cysteines may be unpaired and could therefore form a disulfide bridge with VCP. To test this, we generated a series of A56 mutants in which each cysteine was mutated to a serine, and we found that mutation of cysteine 162 abrogated VCP cell surface expression. We also tested the ability of other poxvirus complement control proteins to bind to VACV A56. While the smallpox homolog of VCP is able to bind VACV A56, the ectromelia virus (ECTV) VCP homolog is only able to bind the ECTV homolog of A56, indicating that these proteins may have coevolved. Surface expression of poxvirus complement control proteins may have important implications in viral pathogenesis, as a virus that does not express cell surface VCP is attenuated in vivo. This suggests that surface expression of VCP may contribute to poxvirus pathogenesis.
DeHaven
Brian C
BC
University of Pennsylvania School of Medicine, Department of Medicine, Division of Infectious Diseases, 502 Johnson Pavilion, Philadelphia, PA 19104-6073, USA.
Girgis
Natasha M
NM
Xiao
Yuhong
Y
Hudson
Paul N
PN
Olson
Victoria A
VA
Damon
Inger K
IK
Isaacs
Stuart N
SN
D002460:Cell Line; D002462:Cell Membrane / Q000821:virology*; D003545:Cysteine / Q000235:genetics / Q000378:metabolism; D004220:Disulfides; D006801:Humans; D016297:Mutagenesis, Site-Directed; D011212:Poxviridae / Q000472:pathogenicity*; D014764:Viral Proteins / Q000378:metabolism*
D004220:Disulfides; D014764:Viral Proteins; C066459:complement-control protein, Vaccinia virus; D003545:Cysteine