Skip to main content
Nucleic Acids Research logoLink to Nucleic Acids Research
. 1986 May 12;14(9):3627–3640. doi: 10.1093/nar/14.9.3627

Self-cleavage of plus and minus RNA transcripts of avocado sunblotch viroid.

C J Hutchins, P D Rathjen, A C Forster, R H Symons
PMCID: PMC339804  PMID: 3714492

Abstract

Self-cleavage of both plus and minus RNA transcripts of the 247-residue avocado sunblotch viroid (ASBV), prepared from tandem dimeric cDNA clones, occurs specifically at two sites in each transcript to give monomeric plus and minus species. The cleavage reaction occurs both during transcription and on incubation of purified transcripts at pH 8 and 37 degrees C in the presence of magnesium ions to give a 3'-terminal 2',3'-cyclic phosphate and a 5'-terminal hydroxyl group. Although the self-cleavage occurs at different sites in the ASBV molecule for the plus and minus species, very similar secondary structures with high sequence homology can be drawn at each site. The results are considered to provide further evidence that ASBV is replicated in vivo by a rolling circle mechanism involving non-enzymic cleavage of high molecular weight RNA precursors of ASBV.

Full text

PDF
3627

Images in this article

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Barker J. M., McInnes J. L., Murphy P. J., Symons R. H. Dot-blot procedure with [32P]DNA probes for the sensitive detection of avocado sunblotch and other viroids in plants. J Virol Methods. 1985 Feb;10(2):87–98. doi: 10.1016/0166-0934(85)90094-1. [DOI] [PubMed] [Google Scholar]
  2. Branch A. D., Robertson H. D. A replication cycle for viroids and other small infectious RNA's. Science. 1984 Feb 3;223(4635):450–455. doi: 10.1126/science.6197756. [DOI] [PubMed] [Google Scholar]
  3. Branch A. D., Robertson H. D., Greer C., Gegenheimer P., Peebles C., Abelson J. Cell-free circularization of viroid progeny RNA by an RNA ligase from wheat germ. Science. 1982 Sep 17;217(4565):1147–1149. doi: 10.1126/science.217.4565.1147. [DOI] [PubMed] [Google Scholar]
  4. Bruce A. G., Uhlenbeck O. C. Reactions at the termini of tRNA with T4 RNA ligase. Nucleic Acids Res. 1978 Oct;5(10):3665–3677. doi: 10.1093/nar/5.10.3665. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Cech T. R., Zaug A. J., Grabowski P. J. In vitro splicing of the ribosomal RNA precursor of Tetrahymena: involvement of a guanosine nucleotide in the excision of the intervening sequence. Cell. 1981 Dec;27(3 Pt 2):487–496. doi: 10.1016/0092-8674(81)90390-1. [DOI] [PubMed] [Google Scholar]
  6. Diener T. O. Viroids. Adv Virus Res. 1983;28:241–283. doi: 10.1016/s0065-3527(08)60725-3. [DOI] [PubMed] [Google Scholar]
  7. Donis-Keller H., Maxam A. M., Gilbert W. Mapping adenines, guanines, and pyrimidines in RNA. Nucleic Acids Res. 1977 Aug;4(8):2527–2538. doi: 10.1093/nar/4.8.2527. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Furneaux H., Pick L., Hurwitz J. Isolation and characterization of RNA ligase from wheat germ. Proc Natl Acad Sci U S A. 1983 Jul;80(13):3933–3937. doi: 10.1073/pnas.80.13.3933. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Garriga G., Lambowitz A. M. RNA splicing in neurospora mitochondria: self-splicing of a mitochondrial intron in vitro. Cell. 1984 Dec;39(3 Pt 2):631–641. doi: 10.1016/0092-8674(84)90470-7. [DOI] [PubMed] [Google Scholar]
  10. Gegenheimer P., Gabius H. J., Peebles C. L., Abelson J. An RNA ligase from wheat germ which participates in transfer RNA splicing in vitro. J Biol Chem. 1983 Jul 10;258(13):8365–8373. [PubMed] [Google Scholar]
  11. Guerrier-Takada C., Gardiner K., Marsh T., Pace N., Altman S. The RNA moiety of ribonuclease P is the catalytic subunit of the enzyme. Cell. 1983 Dec;35(3 Pt 2):849–857. doi: 10.1016/0092-8674(83)90117-4. [DOI] [PubMed] [Google Scholar]
  12. Haseloff J., Symons R. H. Comparative sequence and structure of viroid-like RNAs of two plant viruses. Nucleic Acids Res. 1982 Jun 25;10(12):3681–3691. doi: 10.1093/nar/10.12.3681. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Ishikawa M., Meshi T., Ohno T., Okada Y., Sano T., Ueda I., Shikata E. A revised replication cycle for viroids: the role of longer than unit length RNA in viroid replication. Mol Gen Genet. 1984;196(3):421–428. doi: 10.1007/BF00436189. [DOI] [PubMed] [Google Scholar]
  14. Kan L. S., Chandrasegaran S., Pulford S. M., Miller P. S. Detection of a guanine X adenine base pair in a decadeoxyribonucleotide by proton magnetic resonance spectroscopy. Proc Natl Acad Sci U S A. 1983 Jul;80(14):4263–4265. doi: 10.1073/pnas.80.14.4263. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Keese P., Symons R. H. Domains in viroids: evidence of intermolecular RNA rearrangements and their contribution to viroid evolution. Proc Natl Acad Sci U S A. 1985 Jul;82(14):4582–4586. doi: 10.1073/pnas.82.14.4582. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Kiberstis P. A., Haseloff J., Zimmern D. 2' phosphomonoester, 3'-5' phosphodiester bond at a unique site in a circular viral RNA. EMBO J. 1985 Mar;4(3):817–822. doi: 10.1002/j.1460-2075.1985.tb03703.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Kikuchi Y., Tyc K., Filipowicz W., Sänger H. L., Gross H. J. Circularization of linear viroid RNA via 2'-phosphomonoester, 3', 5'-phosphodiester bonds by a novel type of RNA ligase from wheat germ and Chlamydomonas. Nucleic Acids Res. 1982 Dec 11;10(23):7521–7529. doi: 10.1093/nar/10.23.7521. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Kruger K., Grabowski P. J., Zaug A. J., Sands J., Gottschling D. E., Cech T. R. Self-splicing RNA: autoexcision and autocyclization of the ribosomal RNA intervening sequence of Tetrahymena. Cell. 1982 Nov;31(1):147–157. doi: 10.1016/0092-8674(82)90414-7. [DOI] [PubMed] [Google Scholar]
  19. Krupp G., Gross H. J. Rapid RNA sequencing: nucleases from Staphylococcus aureus and Neurospora crassa discriminate between uridine and cytidine. Nucleic Acids Res. 1979 Aug 10;6(11):3481–3490. doi: 10.1093/nar/6.11.3481. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Melton D. A., Krieg P. A., Rebagliati M. R., Maniatis T., Zinn K., Green M. R. Efficient in vitro synthesis of biologically active RNA and RNA hybridization probes from plasmids containing a bacteriophage SP6 promoter. Nucleic Acids Res. 1984 Sep 25;12(18):7035–7056. doi: 10.1093/nar/12.18.7035. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Owens R. A., Diener T. O. RNA intermediates in potato spindle tuber viroid replication. Proc Natl Acad Sci U S A. 1982 Jan;79(1):113–117. doi: 10.1073/pnas.79.1.113. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Riesner D., Gross H. J. Viroids. Annu Rev Biochem. 1985;54:531–564. doi: 10.1146/annurev.bi.54.070185.002531. [DOI] [PubMed] [Google Scholar]
  23. Sanger F., Coulson A. R., Barrell B. G., Smith A. J., Roe B. A. Cloning in single-stranded bacteriophage as an aid to rapid DNA sequencing. J Mol Biol. 1980 Oct 25;143(2):161–178. doi: 10.1016/0022-2836(80)90196-5. [DOI] [PubMed] [Google Scholar]
  24. Steger G., Hofmann H., Förtsch J., Gross H. J., Randles J. W., Sänger H. L., Riesner D. Conformational transitions in viroids and virusoids: comparison of results from energy minimization algorithm and from experimental data. J Biomol Struct Dyn. 1984 Dec;2(3):543–571. doi: 10.1080/07391102.1984.10507591. [DOI] [PubMed] [Google Scholar]
  25. Symons R. H. Avocado sunblotch viroid: primary sequence and proposed secondary structure. Nucleic Acids Res. 1981 Dec 11;9(23):6527–6537. doi: 10.1093/nar/9.23.6527. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Traub W., Sussman J. L. Adenine-guanine base pairing ribosomal RNA. Nucleic Acids Res. 1982 Apr 24;10(8):2701–2708. doi: 10.1093/nar/10.8.2701. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Visvader J. E., Forster A. C., Symons R. H. Infectivity and in vitro mutagenesis of monomeric cDNA clones of citrus exocortis viroid indicates the site of processing of viroid precursors. Nucleic Acids Res. 1985 Aug 26;13(16):5843–5856. doi: 10.1093/nar/13.16.5843. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Watson N., Gurevitz M., Ford J., Apirion D. Self cleavage of a precursor RNA from bacteriophage T4. J Mol Biol. 1984 Jan 25;172(3):301–323. doi: 10.1016/s0022-2836(84)80028-5. [DOI] [PubMed] [Google Scholar]
  29. Zaug A. J., Grabowski P. J., Cech T. R. Autocatalytic cyclization of an excised intervening sequence RNA is a cleavage-ligation reaction. Nature. 1983 Feb 17;301(5901):578–583. doi: 10.1038/301578a0. [DOI] [PubMed] [Google Scholar]
  30. van der Horst G., Tabak H. F. Self-splicing of yeast mitochondrial ribosomal and messenger RNA precursors. Cell. 1985 Apr;40(4):759–766. doi: 10.1016/0092-8674(85)90335-6. [DOI] [PubMed] [Google Scholar]

Articles from Nucleic Acids Research are provided here courtesy of Oxford University Press

RESOURCES