Detection of virus-specific RNA-dependent RNA polymerase activity in extracts from cells infected with lymphocytic choriomeningitis virus: in vitro synthesis of full-length viral RNA species.
We have developed an in vitro assay for the lymphocytic choriomeningitis virus (LCMV) RNA-dependent RNA polymerase with ribonucleoprotein complexes extracted from acutely infected tissue culture cells. The RNA products synthesized in vitro corresponded in size to the full-length genomic L and S RNAs and subgenomic NP and GP mRNAs normally produced in vivo during acute LCMV infection. In a temporal analysis spanning the first 72 h of acute infection, the in vitro polymerase activity of ribonucleoprotein complexes was maximal at 16 h and declined significantly at later times. In contrast, the intracellular levels of the viral L protein (the putative polymerase protein) appeared to be maximal at 48 to 72 h postinfection. Our results suggest that the accumulation of L protein correlates with reduced viral replication and transcription at later times in acute infection and may be involved in the transition from acute to persistent LCMV infection.
Fuller-Pace
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FV
Department of Immunology, Scripps Clinic and Research Foundation, La Jolla, California 92037.
Southern
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PJ
D000818:Animals; D015153:Blotting, Western; D002460:Cell Line; D002474:Cell-Free System; D006224:Cricetinae; D008216:Lymphocytic Choriomeningitis / Q000201:enzymology / Q000382:microbiology; D008217:Lymphocytic choriomeningitis virus / Q000235:genetics*; D012316:RNA Nucleotidyltransferases / Q000378:metabolism*; D012367:RNA, Viral / Q000096:biosynthesis*; D012324:RNA-Dependent RNA Polymerase / Q000378:metabolism*; D012261:Ribonucleoproteins / Q000378:metabolism; D013997:Time Factors; D014764:Viral Proteins / Q000378:metabolism
D012367:RNA, Viral; D012261:Ribonucleoproteins; D014764:Viral Proteins; D012316:RNA Nucleotidyltransferases; D012324:RNA-Dependent RNA Polymerase