Capsid intermediates assembled in a foot-and-mouth disease virus genome RNA-programmed cell-free translation system and in infected cells.
Structural protein complexes sedimenting at 140S, 70S (empty capsids), and 14S were isolated from foot-and-mouth disease virus-infected cells. The empty capsids were stable, while 14S complexes were relatively short-lived. Radioimmune binding assays involving the use of neutralizing monoclonal antibodies to six distinct epitopes on type A12 virus and polyclonal antisera to A12 structural proteins demonstrated that native empty capsids were indistinguishable from virus. Infected cell 14S particles possessed all the neutralizing epitopes and reacted with VP2 antiserum. Cell-free structural protein complexes sedimenting at 110S, 60S, and 14S containing capsid proteins VP0, VP3, and VP1 are assembled in a rabbit reticulocyte lysate programmed with foot-and-mouth viral RNA. These structures also contain the six epitopes, and cell-free 14S structures like their in vivo counterparts reacted with VP2 antiserum. Capsid structures from infected cells and the cell-free complexes adsorbed to susceptible cells, and this binding was inhibited, to various degrees, by saturating levels of unlabeled virus. These assays and other biochemical evidence indicate that capsid assembly in the cell-free system resembles viral morphogenesis in infected cells. In addition, epitopes on the virus surface possibly involved in interaction with cellular receptor sites are found early in virion morphogenesis.
M J
MJ
Grubman
D O
DO
Morgan
J
J
Kendall
B
B
Baxt
D000818:Animals; D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral / Q000032:analysis; D005537:Aphthovirus / Q000235:genetics / Q000276:immunology / Q000648:ultrastructure*; D002213:Capsid / Q000235:genetics / Q000276:immunology / Q000648:ultrastructure*; D002474:Cell-Free System; D000939:Epitopes; D046911:Macromolecular Substances; D009024:Morphogenesis; D014176:Protein Biosynthesis; D012367:RNA, Viral / Q000235:genetics; D011817:Rabbits; D011991:Receptors, Virus / Q000378:metabolism; D014764:Viral Proteins / Q000235:genetics / Q000302:isolation & purification
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D000939:Epitopes; D046911:Macromolecular Substances; D012367:RNA, Viral; D011991:Receptors, Virus; D014764:Viral Proteins