title: Membrane association of African horsesickness virus nonstructural protein NS3 determines its cytotoxicity.
abstract: The smallest RNA genome segment of African horsesickness virus (AHSV) encodes the nonstructural protein NS3 (24K). NS3 localizes in areas of plasma membrane disruption and is associated with events of viral release. Conserved features in all AHSV NS3 proteins include the synthesis of a truncated NS3A protein from the same open reading frame as that of NS3, a proline-rich region, a region of strict sequence conservation and two hydrophobic domains. To investigate whether these features are associated with the cytotoxicity of NS3 or altered membrane permeability, a series of mutants were constructed and expressed in the BAC-TO-BAC baculovirus-expression system. Our results indicate that mutations in either of the two hydrophobic domains do not prevent membrane targeting of the mutant proteins but abolish their membrane anchoring. This prevents their localization to the cell surface and obviates their cytotoxic effect. The cytotoxicity of NS3 is therefore dependent on its membrane topography and thus involves both hydrophobic domains. NS3 has many of the characteristics of lytic viral proteins that play a central role in viral pathogenesis through modifying membrane permeability.
authors.affiliation: Department of Genetics, University of Pretoria, Pretoria, 0002, South Africa.
authors.forename: M
authors.initials: M
authors.lastname: van Niekerk
authors.forename: C C
authors.initials: CC
authors.lastname: Smit
authors.forename: W C
authors.initials: WC
authors.lastname: Fick
authors.forename: V
authors.initials: V
authors.lastname: van Staden
authors.forename: H
authors.initials: H
authors.lastname: Huismans
mesh_terms: D000356:African Horse Sickness Virus / Q000472:pathogenicity*; D000818:Animals; D016367:Baculoviridae / Q000235:genetics; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D002470:Cell Survival; D005455:Fluorescent Antibody Technique; D009154:Mutation; D012333:RNA, Messenger / Q000032:analysis; D012367:RNA, Viral / Q000032:analysis; D011994:Recombinant Proteins / Q000096:biosynthesis; D018411:Spodoptera; D017361:Viral Nonstructural Proteins / Q000096:biosynthesis / Q000378:metabolism*
chemical_list: C115108:NS3 protein, African horse sickness virus; D012333:RNA, Messenger; D012367:RNA, Viral; D011994:Recombinant Proteins; D017361:Viral Nonstructural Proteins