Skip to main content
PMC home page
The EMBO Journal logoLink to The EMBO Journal
. 1983;2(4):605–610. doi: 10.1002/j.1460-2075.1983.tb01470.x

Ligation of endogenous tRNA 3' half molecules to their corresponding 5' halves via 2'-phosphomonoester,3',5'-phosphodiester bonds in extracts of Chlamydomonas1

Kazimierz Tyc 1,2, Yo Kikuchi 1,3, Maria Konarska 1,2, Witold Filipowicz 1,2,4, Hans J Gross 1,*
PMCID: PMC555068  PMID: 16453445

Abstract

tRNA preparations from Chlamydomonas and wheat germ contain small amounts of tRNA 5' halves and corresponding 3' halves. Incubation of cell-free extracts from the two sources with [γ-32P]ATP yielded 5'-32P-labeled tRNA 3' halves which were joined to their corresponding 5' counterparts to form mature tRNA containing 2'-phosphomonoester,3', 5'-phosphodiester bonds. tRNA 3' halves labelled with T4 kinase were purified, sequenced and also joined to their 5' counterparts. It is proposed that these tRNA halves may be intermediates of the tRNA splicing process, and that the RNA kinase and ligase activities observed here are part of the tRNA splicing complex.

Keywords: plants; endogenous tRNA halves; tRNA ligation; tRNA splicing; 2'-phosphomonoester,3',5'-phosphodiester bonds

Full text

PDF
605

Images in this article

606Fig. 1.
on p.606
606Fig. 2.
on p.606
607Fig. 3.
on p.607
608Fig. 5.
on p.608
608Fig. 6.
on p.608
608Fig. 7.
on p.608

Selected References

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

  1. Abelson J. RNA processing and the intervening sequence problem. Annu Rev Biochem. 1979;48:1035–1069. doi: 10.1146/annurev.bi.48.070179.005131. [DOI] [PubMed] [Google Scholar]
  2. 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]
  3. Burke J. M., RajBhandary U. L. Intron within the large rRNA gene of N. crassa mitochondria: a long open reading frame and a consensus sequence possibly important in splicing. Cell. 1982 Dec;31(3 Pt 2):509–520. doi: 10.1016/0092-8674(82)90307-5. [DOI] [PubMed] [Google Scholar]
  4. Davies R. W., Waring R. B., Ray J. A., Brown T. A., Scazzocchio C. Making ends meet: a model for RNA splicing in fungal mitochondria. Nature. 1982 Dec 23;300(5894):719–724. doi: 10.1038/300719a0. [DOI] [PubMed] [Google Scholar]
  5. De Robertis E. M., Black P., Nishikura K. Intranuclear location of the tRNA splicing enzymes. Cell. 1981 Jan;23(1):89–93. doi: 10.1016/0092-8674(81)90273-7. [DOI] [PubMed] [Google Scholar]
  6. Filipowicz W., Shatkin A. J. Origin of splice junction phosphate in tRNAs processed by HeLa cell extract. Cell. 1983 Feb;32(2):547–557. doi: 10.1016/0092-8674(83)90474-9. [DOI] [PubMed] [Google Scholar]
  7. Gillum A. M., Urquhart N., Smith M., RajBhandary U. L. Nucleotide sequence of salmon testes and salmon liver cytoplasmic initiator tRNA. Cell. 1975 Nov;6(3):395–405. doi: 10.1016/0092-8674(75)90189-0. [DOI] [PubMed] [Google Scholar]
  8. Greer C. L., Peebles C. L., Gegenheimer P., Abelson J. Mechanism of action of a yeast RNA ligase in tRNA splicing. Cell. 1983 Feb;32(2):537–546. doi: 10.1016/0092-8674(83)90473-7. [DOI] [PubMed] [Google Scholar]
  9. Gross H. J., Krupp G., Domdey H., Raba M., Jank P., Lossow C., Alberty H., Ramm K., Sänger H. L. Nucleotide sequence and secondary structure of citrus exocortis and chrysanthemum stunt viroid. Eur J Biochem. 1982 Jan;121(2):249–257. doi: 10.1111/j.1432-1033.1982.tb05779.x. [DOI] [PubMed] [Google Scholar]
  10. Jay E., Bambara R., Padmanabhan R., Wu R. DNA sequence analysis: a general, simple and rapid method for sequencing large oligodeoxyribonucleotide fragments by mapping. Nucleic Acids Res. 1974 Mar;1(3):331–353. doi: 10.1093/nar/1.3.331. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. 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]
  12. Konarska M., Filipowicz W., Domdey H., Gross H. J. Formation of a 2'-phosphomonoester, 3',5'-phosphodiester linkage by a novel RNA ligase in wheat germ. Nature. 1981 Sep 10;293(5828):112–116. doi: 10.1038/293112a0. [DOI] [PubMed] [Google Scholar]
  13. Konarska M., Filipowicz W., Gross H. J. RNA ligation via 2'-phosphomonoester, 3'5'-phosphodiester linkage: requirement of 2',3'-cyclic phosphate termini and involvement of a 5'-hydroxyl polynucleotide kinase. Proc Natl Acad Sci U S A. 1982 Mar;79(5):1474–1478. doi: 10.1073/pnas.79.5.1474. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. 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]
  15. 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]
  16. Lockard R. E., Alzner-Deweerd B., Heckman J. E., MacGee J., Tabor M. W., RajBhandary U. L. Sequence analysis of 5'[32P] labeled mRNA and tRNA using polyacrylamide gel electrophoresis. Nucleic Acids Res. 1978 Jan;5(1):37–56. doi: 10.1093/nar/5.1.37. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Nishikura K., De Robertis E. M. RNA processing in microinjected Xenopus oocytes. Sequential addition of base modifications in the spliced transfer RNA. J Mol Biol. 1981 Jan 15;145(2):405–420. doi: 10.1016/0022-2836(81)90212-6. [DOI] [PubMed] [Google Scholar]
  18. Otsuka A., de Paolis A., Tocchini-Valentini G. P. Ribonuclease "XlaI," an activity from Xenopus laevis oocytes that excises intervening sequences from yeast transfer ribonucleic acid precursors. Mol Cell Biol. 1981 Mar;1(3):269–280. doi: 10.1128/mcb.1.3.269. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Peebles C. L., Ogden R. C., Knapp G., Abelson J. Splicing of yeast tRNA precursors: a two-stage reaction. Cell. 1979 Sep;18(1):27–35. doi: 10.1016/0092-8674(79)90350-7. [DOI] [PubMed] [Google Scholar]
  20. Sharp P. A. Speculations on RNA splicing. Cell. 1981 Mar;23(3):643–646. doi: 10.1016/0092-8674(81)90425-6. [DOI] [PubMed] [Google Scholar]
  21. Simsek M., Ziegenmeyer J., Heckman J., Rajbhandary U. L. Absence of the sequence G-T-psi-C-G(A)- in several eukaryotic cytoplasmic initiator transfer RNAs. Proc Natl Acad Sci U S A. 1973 Apr;70(4):1041–1045. doi: 10.1073/pnas.70.4.1041. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Stackebrandt E., Ludwig W., Schleifer K. H., Gross H. J. Rapid cataloging of ribonuclease T1 resistant oligonucleotides from ribosomal RNAs for phylogenetic studies. J Mol Evol. 1981;17(4):227–236. doi: 10.1007/BF01732760. [DOI] [PubMed] [Google Scholar]
  23. Steinmetz A., Gubbins E. J., Bogorad L. The anticodon of the maize chloroplast gene for tRNA Leu UAA is split by a large intron. Nucleic Acids Res. 1982 May 25;10(10):3027–3037. doi: 10.1093/nar/10.10.3027. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Walker R. T., RajBhandary U. L. The nucleotide sequence of formylmethionine tRNA from Mycoplasma mycoides sp. capri. Nucleic Acids Res. 1978 Jan;5(1):57–70. doi: 10.1093/nar/5.1.57. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Wengler G., Wengler G., Gross H. S. Replicative form of Semliki Forest virus RNA contains an unpaired guanosine. Nature. 1979 Dec 13;282(5740):754–756. doi: 10.1038/282754a0. [DOI] [PubMed] [Google Scholar]
  26. Winicov I., Button J. D. Nuclear ligation of RNA 5'-OH kinase products in tRNA. Mol Cell Biol. 1982 Mar;2(3):241–249. doi: 10.1128/mcb.2.3.241. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from The EMBO Journal are provided here courtesy of Nature Publishing Group

RESOURCES