RNA dependent RNA polymerase activity associated with the double-stranded RNA virus of Giardia lamblia (original) (raw)

Abstract

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.

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  1. Bellofatto V., Cross G. A. Expression of a bacterial gene in a trypanosomatid protozoan. Science. 1989 Jun 9;244(4909):1167–1169. doi: 10.1126/science.2499047. [DOI] [PubMed] [Google Scholar]
  2. Bruenn J. A. Virus-like particles of yeast. Annu Rev Microbiol. 1980;34:49–68. doi: 10.1146/annurev.mi.34.100180.000405. [DOI] [PubMed] [Google Scholar]
  3. Bruenn J., Bobek L., Brennan V., Held W. Yeast viral RNA polymerase is a transcriptase. Nucleic Acids Res. 1980 Jul 11;8(13):2985–2997. doi: 10.1093/nar/8.13.2985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Buck K. W., Ackermann H. W., Bozarth R. F., Bruenn J. A., Koltin Y., Rawlinson C. J., Ushiyama R., Wood H. A. Six groups of double-stranded RNA mycoviruses. Intervirology. 1984;22(1):17–23. doi: 10.1159/000149529. [DOI] [PubMed] [Google Scholar]
  5. Diamond M. E., Dowhanick J. J., Nemeroff M. E., Pietras D. F., Tu C. L., Bruenn J. A. Overlapping genes in a yeast double-stranded RNA virus. J Virol. 1989 Sep;63(9):3983–3990. doi: 10.1128/jvi.63.9.3983-3990.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Fujimura T., Wickner R. B. Gene overlap results in a viral protein having an RNA binding domain and a major coat protein domain. Cell. 1988 Nov 18;55(4):663–671. doi: 10.1016/0092-8674(88)90225-5. [DOI] [PubMed] [Google Scholar]
  7. Fujimura T., Wickner R. B. Reconstitution of template-dependent in vitro transcriptase activity of a yeast double-stranded RNA virus. J Biol Chem. 1989 Jun 25;264(18):10872–10877. [PubMed] [Google Scholar]
  8. Fujimura T., Wickner R. B. Replicase of L-A virus-like particles of Saccharomyces cerevisiae. In vitro conversion of exogenous L-A and M1 single-stranded RNAs to double-stranded form. J Biol Chem. 1988 Jan 5;263(1):454–460. [PubMed] [Google Scholar]
  9. Furfine E. S., White T. C., Wang A. L., Wang C. C. A single-stranded RNA copy of the Giardia lamblia virus double-stranded RNA genome is present in the infected Giardia lamblia. Nucleic Acids Res. 1989 Sep 25;17(18):7453–7467. doi: 10.1093/nar/17.18.7453. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Groot G. S., Poyton R. O. Oxygen control of cytochrome c oxidase synthesis in isolated mitochondria from Saccharomyces cerevisiae. Nature. 1975 May 15;255(5505):238–240. doi: 10.1038/255238a0. [DOI] [PubMed] [Google Scholar]
  11. Hastie N. D., Brennan V., Bruenn J. A. No homology between double-stranded RNA and nuclear DNA of yeast. J Virol. 1978 Dec;28(3):1002–1005. doi: 10.1128/jvi.28.3.1002-1005.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Herring A. J., Bevan E. A. Yeast virus-like particles possess a capsid-associated single-stranded RNA polymerase. Nature. 1977 Aug 4;268(5619):464–466. doi: 10.1038/268464a0. [DOI] [PubMed] [Google Scholar]
  13. Holland M. J., Hager G. L., Rutter W. J. Characterization of purified poly(adenylic acid)-containing messenger ribonucleic acid from Saccharomyces cerevisiae. Biochemistry. 1977 Jan 11;16(1):8–16. doi: 10.1021/bi00620a002. [DOI] [PubMed] [Google Scholar]
  14. Icho T., Wickner R. B. The double-stranded RNA genome of yeast virus L-A encodes its own putative RNA polymerase by fusing two open reading frames. J Biol Chem. 1989 Apr 25;264(12):6716–6723. [PubMed] [Google Scholar]
  15. Miller R. L., Wang A. L., Wang C. C. Identification of Giardia lamblia isolates susceptible and resistant to infection by the double-stranded RNA virus. Exp Parasitol. 1988 Jun;66(1):118–123. doi: 10.1016/0014-4894(88)90056-2. [DOI] [PubMed] [Google Scholar]
  16. Miller R. L., Wang A. L., Wang C. C. Purification and characterization of the Giardia lamblia double-stranded RNA virus. Mol Biochem Parasitol. 1988 Apr;28(3):189–195. doi: 10.1016/0166-6851(88)90003-5. [DOI] [PubMed] [Google Scholar]
  17. Nemeroff M. E., Bruenn J. A. Conservative replication and transcription of Saccharomyces cerevisiae viral double-stranded RNA in vitro. J Virol. 1986 Mar;57(3):754–758. doi: 10.1128/jvi.57.3.754-758.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Pickering L. K., Engelkirk P. G. Giardia lamblia. Pediatr Clin North Am. 1988 Jun;35(3):565–577. doi: 10.1016/s0031-3955(16)36472-0. [DOI] [PubMed] [Google Scholar]
  19. Reich E., Goldberg I. H. Actinomycin and nucleic acid function. Prog Nucleic Acid Res Mol Biol. 1964;3:183–234. doi: 10.1016/s0079-6603(08)60742-4. [DOI] [PubMed] [Google Scholar]
  20. Silverstein S. C., Christman J. K., Acs G. The reovirus replicative cycle. Annu Rev Biochem. 1976;45:375–408. doi: 10.1146/annurev.bi.45.070176.002111. [DOI] [PubMed] [Google Scholar]
  21. Wang A. L., Wang C. C. Discovery of a specific double-stranded RNA virus in Giardia lamblia. Mol Biochem Parasitol. 1986 Dec;21(3):269–276. doi: 10.1016/0166-6851(86)90132-5. [DOI] [PubMed] [Google Scholar]
  22. Wehrli W., Knüsel F., Schmid K., Staehelin M. Interaction of rifamycin with bacterial RNA polymerase. Proc Natl Acad Sci U S A. 1968 Oct;61(2):667–673. doi: 10.1073/pnas.61.2.667. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Welsh D., Leibowitz M. J. Transcription of killer virion double-stranded RNA in vitro. Nucleic Acids Res. 1980 Jun 11;8(11):2365–2375. doi: 10.1093/nar/8.11.2365. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Welsh J. D., Leibowitz M. J., Wickner R. B. Virion DNA-independent RNA polymerase from Saccharomyces cerevisiae. Nucleic Acids Res. 1980 Jun 11;8(11):2349–2363. doi: 10.1093/nar/8.11.2349. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Wickner R. B. Double-stranded RNA replication in yeast: the killer system. Annu Rev Biochem. 1986;55:373–395. doi: 10.1146/annurev.bi.55.070186.002105. [DOI] [PubMed] [Google Scholar]