Foot-and-mouth disease virus particles contain replicase protein 3D.
An antibody against the Escherichia coli-expressed RNA polymerase of foot-and-mouth disease virus (FMDV) reacts with the virus in ELISA and radioimmunoprecipitation experiments and with a protein of the disrupted virus particle in an immunoblot analysis. Treatment of the virus with trypsin, which cleaves capsid protein VP1 and a 56-kDa polypeptide present in trace amount in the particles, reduces the level of the reaction in ELISA and radioimmunoprecipitation and eliminates the immunoblot reaction. Electron microscopy showed that only approximately 20% of the virus particles reacted with the anti-polymerase antibody, whereas most reacted with an antibody against the immunodominant G-H loop of the virus. In the presence of ammonium ions, the expressed polymerase degrades the RNA of the virus into molecules sedimenting at approximately 12 S, indicating that it can act as a hydrolytic as well as a polymerizing enzyme. Moreover, the RNA in trypsin-treated virus particles is degraded when incubated at 37 degrees C, suggesting that the cleaved 56-kDa protein still possesses hydrolytic activity. In addition, the anti-polymerase antibody, which inhibits the polymerase activity of the E. coli-expressed protein, also partially inhibits the hydrolytic activity of the previously described endonuclease of the virus particle, suggesting that this enzyme is identical with the polymerase or forms part of it.
Newman
J F
JF
U.S. Department of Agriculture, Plum Island Animal Disease Center, Greenport, NY 11944.
Piatti
P G
PG
Gorman
B M
BM
Burrage
T G
TG
Ryan
M D
MD
Flint
M
M
Brown
F
F
D000914:Antibodies, Viral; D005537:Aphthovirus / Q000201:enzymology* / Q000235:genetics / Q000276:immunology; D001483:Base Sequence; D004279:DNA, Viral / Q000235:genetics; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000235:genetics; D008854:Microscopy, Electron; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D011233:Precipitin Tests; D012324:RNA-Dependent RNA Polymerase / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D014764:Viral Proteins / Q000276:immunology / Q000302:isolation & purification
D000914:Antibodies, Viral; D004279:DNA, Viral; D014764:Viral Proteins; D012324:RNA-Dependent RNA Polymerase