title: Insertional inactivation of the vaccinia virus 32-kilodalton gene is associated with attenuation in mice and reduction of viral gene expression in polarized epithelial cells.
abstract: The mechanism of poxvirus attachment to cells is poorly understood. We have identified a 32-kDa envelope protein of vaccinia virus which binds to the surface of cultured cells. This binding is specific and selective (J.-S. Maa, J. F. Rodriguez, and M. Esteban, J. Biol. Chem. 265:22174-22180, 1990; C. Lai, S. Gong, and M. Esteban, J. Virol. 65:499-504, 1991). In this investigation, we studied the effect of inactivating the 32-kDa gene (32K gene) on the biology of vaccinia virus. We show that inactivation of the 32K gene decreases by 80% the mortality of mice infected with 32K- vaccinia virus. This reduction in mortality correlates with diminished viral gene expression in target tissues. In highly polarized epithelial cells, viral gene expression of 32K- virus was reduced (50 to 60%) at both the apical and basolateral surfaces in comparison with a 32K+ virus. Restriction of virus gene expression in polarized cell surfaces occurs for both intracellular and extracellular forms of infectious 32K- vaccinia virus. The two infectious forms of vaccinia virus 32K+ infect polarized cells preferentially by the basolateral surface. Our findings provide evidence of the importance of the 32-kDa protein in viral pathogenesis.
authors.affiliation: Department of Biochemistry, State University of New York Health Science Center, Brooklyn 11203-2098.
authors.forename: J R
authors.initials: JR
authors.lastname: Rodriguez
authors.forename: D
authors.initials: D
authors.lastname: Rodriguez
authors.forename: M
authors.initials: M
authors.lastname: Esteban
mesh_terms: D000818:Animals; D002460:Cell Line; D004847:Epithelial Cells; D005260:Female; D015967:Gene Expression Regulation, Viral; D008156:Luciferases / Q000235:genetics; D051379:Mice; D008807:Mice, Inbred BALB C; D014613:Vaccines, Attenuated; D014616:Vaccinia virus / Q000235:genetics / Q000472:pathogenicity / Q000502:physiology*; D014759:Viral Envelope Proteins / Q000235:genetics / Q000378:metabolism*; D014779:Virus Replication
chemical_list: C072001:32kDa protein, Vaccinia virus; D014613:Vaccines, Attenuated; D014759:Viral Envelope Proteins; D008156:Luciferases