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. 1995 Sep 12;92(19):8994–8998. doi: 10.1073/pnas.92.19.8994

Single-site cleavage in the 5'-untranslated region of Leishmaniavirus RNA is mediated by the viral capsid protein.

K J MacBeth 1, J L Patterson 1
PMCID: PMC41094  PMID: 7568059

Abstract

Leishmaniavirus (LRV) is a double-stranded RNA virus that persistently infects the protozoan parasite Leishmania. LRV produces a short RNA transcript, corresponding to the 5' end of positive-sense viral RNA, both in vivo and in in vitro polymerase assays. The short transcript is generated by a single site-specific cleavage event in the 5' untranslated region of the 5.3-kb genome. This cleavage event can be reproduced in vitro with purified viral particles and a substrate RNA transcript possessing the viral cleavage site. A region of nucleotides required for cleavage was identified by analyzing the cleavage sites yielding the short transcripts of various LRV isolates. A 6-nt deletion at this cleavage site completely abolished RNA processing. In an in vitro cleavage assay, baculovirus-expressed capsid protein possessed an endonuclease activity identical to that of native virions, showing that the viral capsid protein is the RNA endonuclease. Identification of the LRV capsid protein as an RNA endonuclease is unprecedented among known viral capsid proteins.

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Selected References

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  1. Armstrong T. C., Patterson J. L. Cultivation of Leishmania braziliensis in an economical serum-free medium containing human urine. J Parasitol. 1994 Dec;80(6):1030–1032. [PubMed] [Google Scholar]
  2. Bates P. A. Characterization of developmentally-regulated nucleases in promastigotes and amastigotes of Leishmania mexicana. FEMS Microbiol Lett. 1993 Feb 15;107(1):53–58. doi: 10.1016/0378-1097(93)90353-4. [DOI] [PubMed] [Google Scholar]
  3. Bruenn J. A. Relationships among the positive strand and double-strand RNA viruses as viewed through their RNA-dependent RNA polymerases. Nucleic Acids Res. 1991 Jan 25;19(2):217–226. doi: 10.1093/nar/19.2.217. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Cadd T. L., Keenan M. C., Patterson J. L. Detection of Leishmania RNA virus 1 proteins. J Virol. 1993 Sep;67(9):5647–5650. doi: 10.1128/jvi.67.9.5647-5650.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Cadd T. L., MacBeth K., Furlong D., Patterson J. L. Mutational analysis of the capsid protein of Leishmania RNA virus LRV1-4. J Virol. 1994 Dec;68(12):7738–7745. doi: 10.1128/jvi.68.12.7738-7745.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Cadd T. L., Patterson J. L. Synthesis of viruslike particles by expression of the putative capsid protein of Leishmania RNA virus in a recombinant baculovirus expression system. J Virol. 1994 Jan;68(1):358–365. doi: 10.1128/jvi.68.1.358-365.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Campbell T. A., Zlotnick G. W., Neubert T. A., Sacci J. B., Jr, Gottlieb M. Purification and characterization of the 3'-nucleotidase/nuclease from promastigotes of Leishmania donovani. Mol Biochem Parasitol. 1991 Jul;47(1):109–117. doi: 10.1016/0166-6851(91)90153-w. [DOI] [PubMed] [Google Scholar]
  8. Choi G. H., Nuss D. L. Hypovirulence of chestnut blight fungus conferred by an infectious viral cDNA. Science. 1992 Aug 7;257(5071):800–803. doi: 10.1126/science.1496400. [DOI] [PubMed] [Google Scholar]
  9. Chung I. K., Armstrong T. C., Patterson J. L. Identification of a short viral transcript in Leishmania RNA virus-infected cells. Virology. 1994 Feb;198(2):552–556. doi: 10.1006/viro.1994.1066. [DOI] [PubMed] [Google Scholar]
  10. Guilbride L., Myler P. J., Stuart K. Distribution and sequence divergence of LRV1 viruses among different Leishmania species. Mol Biochem Parasitol. 1992 Aug;54(1):101–104. doi: 10.1016/0166-6851(92)90099-6. [DOI] [PubMed] [Google Scholar]
  11. Harris M., Decker C., Sollner-Webb B., Hajduk S. Specific cleavage of pre-edited mRNAs in trypanosome mitochondrial extracts. Mol Cell Biol. 1992 Jun;12(6):2591–2598. doi: 10.1128/mcb.12.6.2591. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. MacBeth K. J., Patterson J. L. The short transcript of Leishmania RNA virus is generated by RNA cleavage. J Virol. 1995 Jun;69(6):3458–3464. doi: 10.1128/jvi.69.6.3458-3464.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Patterson J. L. The current status of Leishmania RNA virus I. Parasitol Today. 1993 Apr;9(4):135–136. doi: 10.1016/0169-4758(93)90178-i. [DOI] [PubMed] [Google Scholar]
  14. Scheffter S., Widmer G., Patterson J. L. Complete sequence of Leishmania RNA virus 1-4 and identification of conserved sequences. Virology. 1994 Mar;199(2):479–483. doi: 10.1006/viro.1994.1149. [DOI] [PubMed] [Google Scholar]
  15. Stuart K. D., Weeks R., Guilbride L., Myler P. J. Molecular organization of Leishmania RNA virus 1. Proc Natl Acad Sci U S A. 1992 Sep 15;89(18):8596–8600. doi: 10.1073/pnas.89.18.8596. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Sutton R. E., Boothroyd J. C. Evidence for trans splicing in trypanosomes. Cell. 1986 Nov 21;47(4):527–535. doi: 10.1016/0092-8674(86)90617-3. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Tarr P. I., Aline R. F., Jr, Smiley B. L., Scholler J., Keithly J., Stuart K. LR1: a candidate RNA virus of Leishmania. Proc Natl Acad Sci U S A. 1988 Dec;85(24):9572–9575. doi: 10.1073/pnas.85.24.9572. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. 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]
  19. Wang A. L., Wang C. C. The double-stranded RNA in Trichomonas vaginalis may originate from virus-like particles. Proc Natl Acad Sci U S A. 1986 Oct;83(20):7956–7960. doi: 10.1073/pnas.83.20.7956. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Widmer G., Comeau A. M., Furlong D. B., Wirth D. F., Patterson J. L. Characterization of a RNA virus from the parasite Leishmania. Proc Natl Acad Sci U S A. 1989 Aug;86(15):5979–5982. doi: 10.1073/pnas.86.15.5979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Widmer G., Patterson J. L. Genomic structure and RNA polymerase activity in Leishmania virus. J Virol. 1991 Aug;65(8):4211–4215. doi: 10.1128/jvi.65.8.4211-4215.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]

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