title: Self-assembly and protein-protein interactions between the SV40 capsid proteins produced in insect cells.
abstract: Soluble SV40 capsid proteins were obtained by expression of the three late genes, VP1, VP2, and VP3, in Sf9 cells using baculovirus expression vectors. Coproduction of the capsid proteins VP1, VP2, and VP3 was achieved by infecting Sf9 cells with the three recombinant baculovirus species at equal multiplicities. All three proteins were found to be localized in the nuclear fraction. Electron microscopy of nuclear extracts of the infected cells showed an abundance of SV40-like capsid structures and heterogeneous aggregates of variable size, mostly 20-45 nm. Under the same staining conditions wild-type SV40 virions are 45 nm. The capsid-like particles sedimented in glycerol gradients similarly to authentic wild-type SV40 virions. Pentamers of the major capsid protein VP1 were also seen. Protein analysis on sucrose gradients demonstrated that the capsid-like particles can be disrupted by treatment with the reducing agent dithiothreitol and the calcium chelator EGTA. The capsid-like particles were found to be significantly less stable than SV40 virions and were partially stabilized by calcium ions. Understanding the complex interactions between the capsid proteins is important for the development of an efficient in vitro packaging system for SV40 virions and pseudovirions.
authors.affiliation: Department of Hematology, The Hebrew University-Hadassah Medical School, Jerusalem, 91120, Israel.
authors.forename: Z
authors.initials: Z
authors.lastname: Sandalon
authors.forename: A
authors.initials: A
authors.lastname: Oppenheim
mesh_terms: D000818:Animals; D015153:Blotting, Western; D002213:Capsid / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism* / Q000648:ultrastructure; D002460:Cell Line; D007313:Insecta; D008854:Microscopy, Electron; D011485:Protein Binding; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism / Q000648:ultrastructure; D013539:Simian virus 40 / Q000378:metabolism*
chemical_list: D011994:Recombinant Proteins