title: Preliminary x-ray characterization of authentic providence virus and attempts to express its coat protein gene in recombinant baculovirus.
abstract: Providence Virus (PrV) is a non-envoloped, T = 4 icosahedral beta-tetravirus that undergoes autocatalytic cleavage of its coat protein precursor after capsid assembly. This is also a well characterized function of Nudaurelia capensis omega virus (NomegaV), a member of the related omegatetraviruses, whose x-ray structure has been determined. Virus-like particle (VLP) production of PrV in a recombinant baculovirus expression system was attempted to obtain high VLP yields for comparison of structural and autocatalytic active site properties between these virus groups. This resulted in insoluble aggregates of PrV coat protein even though NomegaV VLPs have been successfully produced in the same system. Betatetraviruses may be more dependent on compartmentalization and availability of their full-length genome for proper folding and assembly. However, crystals were grown of limited quantities of authentic PrV produced in cell culture and a partial X-ray data set collected to 3.8 A resolution. The virus particle position and orientation in the unit cell was determined by space group consideration and rotation function analysis. A phasing model, based on NomegaV, was developed to initiate the structure solution of PrV.
authors.affiliation: Department of Chemistry and Biochemistry, University of California, San Diego, La Jolla, CA 92037, USA.
authors.forename: D J
authors.initials: DJ
authors.lastname: Taylor
authors.forename: J A
authors.initials: JA
authors.lastname: Speir
authors.forename: V
authors.initials: V
authors.lastname: Reddy
authors.forename: G
authors.initials: G
authors.lastname: Cingolani
authors.forename: F M
authors.initials: FM
authors.lastname: Pringle
authors.forename: L A
authors.initials: LA
authors.lastname: Ball
authors.forename: J E
authors.initials: JE
authors.lastname: Johnson
mesh_terms: D016367:Baculoviridae / Q000235:genetics; D036022:Capsid Proteins / Q000235:genetics; D018360:Crystallography, X-Ray; D007304:Insect Viruses / Q000737:chemistry* / Q000648:ultrastructure*; D008854:Microscopy, Electron; D012399:Rotation; D014162:Transfection; D019065:Virus Assembly
chemical_list: D036022:Capsid Proteins