Transcriptase activity associated with rabies virion

Abstract

Rabies virion-associated transcriptase activity was investigated in vitro and and compared with that of the New Jersey serotype of vesicular stomatitis virus. The concentration of detergent that affected [3H]GMP incorporation into acid-insoluble material was significantly different for both viruses. Vesicular stomatitis virus New Jersey required 0.05-0.1% nonionic detergent, but rabies virion could not be fully activated unless 4-5% detergent was used. Other optimal conditions were as follows: 40 mM NaCl, 5 mM Mg2+, 40 mM Tris-HCl (pH 7.4), 5 mM dithiothreitol and 30.degree. C. The reaction required 4 nucleoside triphosphates. The initial rate of RNA synthesis by rabies virion enzyme was 140 pmol of GMP incorporated/mg of viral protein/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.degree. C [in baby hamster kidney BHK-21 cells]. 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.