Toward an in vitro system for picornavirus assembly: purification of mengovirus 14S capsid precursor particles.
Mengovirus 14S subviral protein particles generated in infected L cells and in a cell-free translation system primed with mengovirus RNA were purified by sucrose gradient centrifugation and immunoaffinity chromatography. The preparations from both sources contained essentially pure proteins epsilon, alpha, and gamma, as was demonstrated in terms of virus-specific proteins (by autoradiography) and total protein content (by silver staining of sodium dodecyl sulfate-polyacrylamide electrophoresis gels). These purified proteins sedimented as discrete particles at the 14S position when recentrifuged in sucrose gradients. Although their assembly properties have not yet been studied in detail, preliminary results indicate that during incubation with virion RNA the 14S particles purified from infected cells can form a structure cosedimenting with mature mengovirus.
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000914:Antibodies, Viral / Q000276:immunology; D002213:Capsid / Q000096:biosynthesis / Q000276:immunology / Q000302:isolation & purification*; D002474:Cell-Free System; D002846:Chromatography, Affinity; D007739:L Cells; D008574:Mengovirus / Q000032:analysis / Q000254:growth & development* / Q000276:immunology; D051379:Mice; D009024:Morphogenesis; D014176:Protein Biosynthesis; D011498:Protein Precursors / Q000096:biosynthesis / Q000276:immunology / Q000302:isolation & purification*; D012367:RNA, Viral / Q000502:physiology; D011817:Rabbits; D012156:Reticulocytes; D014771:Virion / Q000648:ultrastructure; D014776:Virus Cultivation
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D011498:Protein Precursors; D012367:RNA, Viral