RNA dependent RNA polymerase activity associated with the double-stranded RNA virus of Giardia lamblia.
Giardia lamblia, a parasitic protozoan, can contain a double-stranded RNA (dsRNA) virus, GLV (1). We have identified an RNA polymerase activity present specifically in cultures of GLV infected cells. This RNA polymerase activity is present in crude whole cell lysates as well as in lysates from GLV particles purified from the culture medium. The RNA polymerase has many characteristics common to other RNA polymerases (e.g. it requires divalent cations and all four ribonucleoside triphosphates), yet it is not inhibited by RNA polymerase inhibitors such as alpha-amanitin or rifampicin. The RNA polymerase activity synthesizes RNAs corresponding to one strand of the GLV genome, although under the present experimental conditions, the RNA products of the reaction are not full length viral RNAs. The in vitro products of the RNA polymerase reaction co-sediment through sucrose gradients with viral particles; and purified GLV viral particles have RNA polymerase activity. The RNA polymerase activities within and outside of infected cells closely parallel the amount of virus present during the course of viral infection. The similarities between the RNA polymerase of GLV and the polymerase associated with the dsRNA virus system of yeast are discussed.
White
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Department of Pharmaceutical Chemistry, School of Pharmacy, University of California, San Francisco 94143-0446.
Wang
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D000818:Animals; D002413:Cations, Divalent; D002499:Centrifugation, Density Gradient; D003570:Cytidine Triphosphate / Q000378:metabolism; D004587:Electrophoresis, Agar Gel; D005872:Giardia / Q000382:microbiology*; D006358:Hot Temperature; D007700:Kinetics; D008274:Magnesium / Q000494:pharmacology; D012316:RNA Nucleotidyltransferases / Q000378:metabolism*; D012328:RNA Viruses / Q000201:enzymology*; D012367:RNA, Viral / Q000096:biosynthesis / Q000302:isolation & purification; D012324:RNA-Dependent RNA Polymerase / Q000037:antagonists & inhibitors / Q000302:isolation & purification / Q000378:metabolism*; D012260:Ribonucleases / Q000494:pharmacology; D012967:Sodium Dodecyl Sulfate / Q000494:pharmacology; D014771:Virion / Q000201:enzymology / Q000302:isolation & purification
D002413:Cations, Divalent; D012367:RNA, Viral; D012967:Sodium Dodecyl Sulfate; D003570:Cytidine Triphosphate; D012316:RNA Nucleotidyltransferases; D012324:RNA-Dependent RNA Polymerase; D012260:Ribonucleases; D008274:Magnesium