Isolation of a foot-and-mouth disease polyuridylic acid polymerase and its inhibition by antibody.
A template-dependent polyuridylic acid [poly(U)] polymerase has been isolated from BHK cells infected with foot-and-mouth disease virus (FMDV). Enzyme activity in a 20,000 x g supernatant of a cytoplasmic extract was concentrated by precipitation with 30 to 50% saturated ammonium sulfate. The poly(U) polymerase was freed of membranes by sodium dodecyl sulfate and 1,1,2-trichlorotrifluoroethane extraction, and RNA was removed by precipitation with 2 M LiCl. The solubilized poly(U) polymerase required polyadenylic acid as template complexed to an oligouridylic acid primer and Mg2+ for activity, but was inhibited by Mn2+. Antisera from animals infected with FMDV had previously been shown to inhibit the activity of FMDV RNA replicase complexed to the endogenous RNA template. The same antisera also inhibited the activity of poly(U) polymerase. Antisera depleted of antibody by absorption with the virus infection-associated antigen of FMDV no longer inhibited replicase and polymerase activities. The evidence suggests that FMDV RNA replicase, poly(U) polymerase, and the virus infection-associated antigen share a common protein.
D000818:Animals; D000914:Antibodies, Viral; D005537:Aphthovirus / Q000201:enzymology*; D002460:Cell Line; D006224:Cricetinae; D006168:Guinea Pigs; D007106:Immune Sera / Q000494:pharmacology; D007668:Kidney; D008647:Mesocricetus; D009713:Nucleotidyltransferases / Q000037:antagonists & inhibitors / Q000276:immunology / Q000302:isolation & purification*; D011061:Poly A; D011072:Poly U / Q000037:antagonists & inhibitors / Q000096:biosynthesis* / Q000276:immunology / Q000302:isolation & purification; D012367:RNA, Viral / Q000096:biosynthesis; D013379:Substrate Specificity; D013698:Templates, Genetic; D014779:Virus Replication
D000914:Antibodies, Viral; D007106:Immune Sera; D012367:RNA, Viral; D011061:Poly A; D011072:Poly U; D009713:Nucleotidyltransferases; C022428:poly U polymerase