Skip to main content
NCBI home page
Nucleic Acids Research logoLink to Nucleic Acids Research
. 1991 Feb 11;19(3):605–609. doi: 10.1093/nar/19.3.605

The rate and specificity of a group I ribozyme are inversely affected by choice of monovalent salt.

S Partono 1, A S Lewin 1
PMCID: PMC333655  PMID: 2011532

Abstract

The fifth intron of the COB gene of yeast mitochondria splices autocatalytically. The rate of splicing is increased by high concentrations of monovalent salts, but the choice of both cation and anion is significant: The smaller the cation in solution, the faster the reaction (the rate in K+ greater than NH4+ greater than Na+ greater than Li+). Chloride, bromide, iodide and acetate salts enhance autocatalytic processing, but sulfate salts do not and fluoride salts are inhibitory. The choice of monovalent salt affects the KM of the intron for guanosine nucleotide, implying an alteration in the affinity of the RNA for that substrate. Under optimal conditions (1M KCl, 50 mM MgCl2) the catalytic efficiency of this intron exceeds that reported for the ribosomal intron from Tetrahymena, but several side reactions occur, including guanosine-addition within the downstream exon. The site of addition resembles the 5' splice junction, but selection of this site does not involve the internal guide sequence of the intron.

Full text

PDF
605

Images in this article

606Image
on p.606
607Image
on p.607
608Image
on p.608

Selected References

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

  1. Bass B. L., Cech T. R. Ribozyme inhibitors: deoxyguanosine and dideoxyguanosine are competitive inhibitors of self-splicing of the Tetrahymena ribosomal ribonucleic acid precursor. Biochemistry. 1986 Aug 12;25(16):4473–4477. doi: 10.1021/bi00364a001. [DOI] [PubMed] [Google Scholar]
  2. Bass B. L., Cech T. R. Specific interaction between the self-splicing RNA of Tetrahymena and its guanosine substrate: implications for biological catalysis by RNA. 1984 Apr 26-May 2Nature. 308(5962):820–826. doi: 10.1038/308820a0. [DOI] [PubMed] [Google Scholar]
  3. Burke J. M. Molecular genetics of group I introns: RNA structures and protein factors required for splicing--a review. Gene. 1988 Dec 20;73(2):273–294. doi: 10.1016/0378-1119(88)90493-3. [DOI] [PubMed] [Google Scholar]
  4. Cech T. R. Conserved sequences and structures of group I introns: building an active site for RNA catalysis--a review. Gene. 1988 Dec 20;73(2):259–271. doi: 10.1016/0378-1119(88)90492-1. [DOI] [PubMed] [Google Scholar]
  5. Collins K. D., Washabaugh M. W. The Hofmeister effect and the behaviour of water at interfaces. Q Rev Biophys. 1985 Nov;18(4):323–422. doi: 10.1017/s0033583500005369. [DOI] [PubMed] [Google Scholar]
  6. Donis-Keller H. Site specific enzymatic cleavage of RNA. Nucleic Acids Res. 1979 Sep 11;7(1):179–192. doi: 10.1093/nar/7.1.179. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Flor P. J., Flanegan J. B., Cech T. R. A conserved base pair within helix P4 of the Tetrahymena ribozyme helps to form the tertiary structure required for self-splicing. EMBO J. 1989 Nov;8(11):3391–3399. doi: 10.1002/j.1460-2075.1989.tb08503.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Gampel A., Nishikimi M., Tzagoloff A. CBP2 protein promotes in vitro excision of a yeast mitochondrial group I intron. Mol Cell Biol. 1989 Dec;9(12):5424–5433. doi: 10.1128/mcb.9.12.5424. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Inoue T., Sullivan F. X., Cech T. R. New reactions of the ribosomal RNA precursor of Tetrahymena and the mechanism of self-splicing. J Mol Biol. 1986 May 5;189(1):143–165. doi: 10.1016/0022-2836(86)90387-6. [DOI] [PubMed] [Google Scholar]
  10. Jarrell K. A., Peebles C. L., Dietrich R. C., Romiti S. L., Perlman P. S. Group II intron self-splicing. Alternative reaction conditions yield novel products. J Biol Chem. 1988 Mar 5;263(7):3432–3439. [PubMed] [Google Scholar]
  11. Lamb M. R., Anziano P. Q., Glaus K. R., Hanson D. K., Klapper H. J., Perlman P. S., Mahler H. R. Functional domains in introns. RNA processing intermediates in cis- and trans-acting mutants in the penultimate intron of the mitochondrial gene for cytochrome b. J Biol Chem. 1983 Feb 10;258(3):1991–1999. [PubMed] [Google Scholar]
  12. McGraw P., Tzagoloff A. Assembly of the mitochondrial membrane system. Characterization of a yeast nuclear gene involved in the processing of the cytochrome b pre-mRNA. J Biol Chem. 1983 Aug 10;258(15):9459–9468. [PubMed] [Google Scholar]
  13. Michel F., Hanna M., Green R., Bartel D. P., Szostak J. W. The guanosine binding site of the Tetrahymena ribozyme. Nature. 1989 Nov 23;342(6248):391–395. doi: 10.1038/342391a0. [DOI] [PubMed] [Google Scholar]
  14. Partono S., Lewin A. S. Autocatalytic activities of intron 5 of the cob gene of yeast mitochondria. Mol Cell Biol. 1988 Jun;8(6):2562–2571. doi: 10.1128/mcb.8.6.2562. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Price J. V., Engberg J., Cech T. R. 5' exon requirement for self-splicing of the Tetrahymena thermophila pre-ribosomal RNA and identification of a cryptic 5' splice site in the 3' exon. J Mol Biol. 1987 Jul 5;196(1):49–60. doi: 10.1016/0022-2836(87)90510-9. [DOI] [PubMed] [Google Scholar]
  16. Reich C., Olsen G. J., Pace B., Pace N. R. Role of the protein moiety of ribonuclease P, a ribonucleoprotein enzyme. Science. 1988 Jan 8;239(4836):178–181. doi: 10.1126/science.3122322. [DOI] [PubMed] [Google Scholar]
  17. Suh E. R., Waring R. B. Base pairing between the 3' exon and an internal guide sequence increases 3' splice site specificity in the Tetrahymena self-splicing rRNA intron. Mol Cell Biol. 1990 Jun;10(6):2960–2965. doi: 10.1128/mcb.10.6.2960. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Waring R. B. Identification of phosphate groups important to self-splicing of the Tetrahymena rRNA intron as determined by phosphorothioate substitution. Nucleic Acids Res. 1989 Dec 25;17(24):10281–10293. doi: 10.1093/nar/17.24.10281. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Zaug A. J., Cech T. R. Oligomerization of intervening sequence RNA molecules in the absence of proteins. Science. 1985 Sep 13;229(4718):1060–1064. doi: 10.1126/science.2412290. [DOI] [PubMed] [Google Scholar]

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

RESOURCES