title: Cell surface expression of the vaccinia virus complement control protein is mediated by interaction with the viral A56 protein and protects infected cells from complement attack.
abstract: The vaccinia virus (VACV) complement control protein (VCP) is the major protein secreted from VACV-infected cells. It has been reported that VCP binds to the surfaces of uninfected cells by interacting with heparan sulfate proteoglycans (HSPGs). In this study, we show that VCP is also expressed on the surfaces of infected cells and demonstrate that surface localization occurs independently of HSPGs. Since VCP does not contain a transmembrane domain, we hypothesized that VCP interacts with a membrane protein that localizes to the infected-cell surface. We show that the VACV A56 membrane protein is necessary for the cell surface expression of VCP and demonstrate that VCP and A56 interact in VACV-infected cells. Since the surface expression of VCP was abrogated by reducing agents, we examined the contribution of an unpaired cysteine residue on VCP to VCP surface expression and VCP's interaction with A56. To do this, we mutated the unpaired cysteine in VCP and generated a recombinant virus expressing the altered form of VCP. Following the infection of cells with the mutant virus, VCP was neither expressed on the cell surface nor able to interact with A56. Importantly, the cell surface expression of VCP was found to protect infected cells from complement-mediated lysis. Our findings suggest a new function for VCP that may be important for poxvirus pathogenesis and impact immune responses to VACV-based vaccines.
authors.affiliation: University of Pennsylvania School of Medicine, Division of Infectious Diseases, 502 Johnson Pavilion, Philadelphia, PA 19104-6073, USA.
authors.forename: Natasha M
authors.initials: NM
authors.lastname: Girgis
authors.forename: Brian C
authors.initials: BC
authors.lastname: Dehaven
authors.forename: Xin
authors.initials: X
authors.lastname: Fan
authors.forename: Kendra M
authors.initials: KM
authors.lastname: Viner
authors.forename: Mohammad
authors.initials: M
authors.lastname: Shamim
authors.forename: Stuart N
authors.initials: SN
authors.lastname: Isaacs
mesh_terms: D002460:Cell Line; D003167:Complement Activation; D003545:Cysteine / Q000235:genetics; D015967:Gene Expression Regulation, Viral / Q000276:immunology; D006493:Heparin / Q000031:analogs & derivatives; D006801:Humans; D016297:Mutagenesis, Site-Directed; D011509:Proteoglycans; D014615:Vaccinia / Q000276:immunology*; D014616:Vaccinia virus / Q000472:pathogenicity*; D017361:Viral Nonstructural Proteins / Q000378:metabolism*; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism* / Q000502:physiology*
chemical_list: D011509:Proteoglycans; D017361:Viral Nonstructural Proteins; D014764:Viral Proteins; C066459:complement-control protein, Vaccinia virus; C036090:heparin proteoglycan; D006493:Heparin; D003545:Cysteine