Transcriptase activity associated with rabies virion.
Rabies virion-associated transcriptase activity was investigated in vitro and compared with that of the New Jersey serotype of vesicular stomatitis virus. The concentration of detergent that affected [3H]GMP incoporation into acid-insoluble material was significantly different for both viruses. Vesicular stomatitis virus New Jersey required 0.05 to 0.1% nonionic detergent, whereas rabies virion could not be fully activated unless 4 to 5% detergent was used. Other optimal conditions were as follows: 40 mM NaCl, 5 mM Mg2+, 40 mM Tris-hydrochloride (pH 7.4), 5 mM dithiothreitol, and 30 degrees C. The reaction required four nucleoside triphosphates. The initial rate of RNA synthesis by rabies virion enzyme was 140 pmol of GMP incorporated/mg of viral protein per h and linearly increased until about 8 h, with a slight initial lag phase. The enzyme activity that correlated with the content of L protein was highest when rabies virions were grown at 33 degrees C. The product was single-stranded RNA, which was complementary in base sequences to rabies viral RNA. Most of the RNA synthesized sedimented at 6-16S.
D012321:DNA-Directed RNA Polymerases / Q000378:metabolism*; D004229:Dithiothreitol / Q000494:pharmacology; D006157:Guanosine Monophosphate / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008274:Magnesium / Q000494:pharmacology; D012367:RNA, Viral / Q000096:biosynthesis; D011820:Rabies virus / Q000201:enzymology*; D018116:Vesiculovirus / Q000201:enzymology; D014771:Virion / Q000201:enzymology*
D012367:RNA, Viral; D006157:Guanosine Monophosphate; D012321:DNA-Directed RNA Polymerases; D008274:Magnesium; D004229:Dithiothreitol