title: The viroporin activity of the minor structural proteins VP2 and VP3 is required for SV40 propagation.
abstract: For nonenveloped viruses such as Simian Virus 40, the mechanism used to translocate viral components across membranes is poorly understood. Previous results indicated that the minor structural proteins, VP2 and VP3, might act as membrane proteins during infection. Here, purified VP2 and VP3 were found to form pores in host cell membranes. To identify possible membrane domains, individual hydrophobic domains from VP2 and VP3 were expressed in a model protein and tested for their ability to integrate into membranes. Several domains from the late proteins supported endoplasmic reticulum membrane insertion as transmembrane domains. Mutations in VP2 and VP3 were engineered that inhibited membrane insertion and pore formation. When these mutations were introduced into the viral genome, viral propagation was inhibited. This comprehensive approach revealed that the viroporin activity of VP2 and VP3 was inhibited by targeted disruptions of individual hydrophobic domains and the loss of membrane disruption activity impaired viral infection.
authors.affiliation: Department of Biochemistry and Molecular Biology, University of Massachusetts, Amherst, Massachusetts 01003, USA.
authors.forename: Kristina M
authors.initials: KM
authors.lastname: Giorda
authors.forename: Smita
authors.initials: S
authors.lastname: Raghava
authors.forename: Macy W
authors.initials: MW
authors.lastname: Zhang
authors.forename: Daniel N
authors.initials: DN
authors.lastname: Hebert
mesh_terms: D000818:Animals; D019556:COS Cells; D036022:Capsid Proteins / Q000502:physiology*; D002522:Chlorocebus aethiops; D016338:Genes, vpu; D016679:Genome, Viral; D005982:Glutathione Transferase / Q000378:metabolism; D054298:Human Immunodeficiency Virus Proteins / Q000378:metabolism*; D008565:Membrane Proteins / Q000737:chemistry; D008954:Models, Biological; D009154:Mutation; D010957:Plasmids / Q000378:metabolism; D015202:Protein Engineering; D017434:Protein Structure, Tertiary; D013539:Simian virus 40 / Q000378:metabolism*; D054334:Viral Regulatory and Accessory Proteins / Q000378:metabolism*; D000086082:Viroporin Proteins
chemical_list: D036022:Capsid Proteins; D005982:Glutathione Transferase; D054298:Human Immunodeficiency Virus Proteins; D008565:Membrane Proteins; D054334:Viral Regulatory and Accessory Proteins; C085902:VP3 protein, Simian virus 40; C514199:Vp2 protein, simian virus 40; C514859:vpu protein, Human immunodeficiency virus 1; D000086082:Viroporin Proteins