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

Nucleic acids research ·Vol. 19 ·No. 3 ·1991-02-11 ·Pages 605-9

Partono S, Lewin AS

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.

Related Genes

CBP2 COB

MeSH Terms

Cations, Monovalent Cytochrome b Group/genetics DNA Mutational Analysis DNA, Mitochondrial/genetics Guanosine Triphosphate/metabolism In Vitro Techniques Introns Kinetics Potassium Chloride/pharmacology RNA Splicing RNA, Catalytic/metabolism RNA, Fungal/metabolism RNA, Messenger/metabolism Saccharomyces cerevisiae/genetics Solvents Substrate Specificity

Chemicals

Cations, Monovalent Cytochrome b Group DNA, Mitochondrial RNA, Catalytic RNA, Fungal RNA, Messenger Solvents Potassium Chloride Guanosine Triphosphate

Authors & Affiliations

2 authors, click to expand affiliations / ORCID

Partono S

Department of Immunology and Medical Microbiology, University of Florida College of Medicine, Gainesville 32610.

Lewin A S

References (19)

19 references, click to expand

Ribozyme inhibitors: deoxyguanosine and dideoxyguanosine are competitive inhibitors of self-splicing of the Tetrahymena ribosomal ribonucleic acid precursor.
Biochemistry. 1986 Aug 12;25(16):4473-7 PMID: 3639741
Oligomerization of intervening sequence RNA molecules in the absence of proteins.
Science. 1985 Sep 13;229(4718):1060-4 PMID: 2412290
The Hofmeister effect and the behaviour of water at interfaces.
Q Rev Biophys. 1985 Nov;18(4):323-422 PMID: 3916340
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 PMID: 2443717
Role of the protein moiety of ribonuclease P, a ribonucleoprotein enzyme.
Science. 1988 Jan 8;239(4836):178-81 PMID: 3122322
Group II intron self-splicing. Alternative reaction conditions yield novel products.
J Biol Chem. 1988 Mar 5;263(7):3432-9 PMID: 2830285
Autocatalytic activities of intron 5 of the cob gene of yeast mitochondria.
Mol Cell Biol. 1988 Jun;8(6):2562-71 PMID: 3043183
Conserved sequences and structures of group I introns: building an active site for RNA catalysis--a review.
Gene. 1988 Dec 20;73(2):259-71 PMID: 3072259
Molecular genetics of group I introns: RNA structures and protein factors required for splicing--a review.
Gene. 1988 Dec 20;73(2):273-94 PMID: 3072260
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):3391-9 PMID: 2684642
CBP2 protein promotes in vitro excision of a yeast mitochondrial group I intron.
Mol Cell Biol. 1989 Dec;9(12):5424-33 PMID: 2685564
The guanosine binding site of the Tetrahymena ribozyme.
Nature. 1989 Nov 23;342(6248):391-5 PMID: 2685606
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):10281-93 PMID: 2690016
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):2960-5 PMID: 2342465
Site specific enzymatic cleavage of RNA.
Nucleic Acids Res. 1979 Sep 11;7(1):179-92 PMID: 386279
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):1991-9 PMID: 6296117
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):9459-68 PMID: 6348045
Specific interaction between the self-splicing RNA of Tetrahymena and its guanosine substrate: implications for biological catalysis by RNA.
Nature. 1984 Apr 26-May 2;308(5962):820-6 PMID: 6562377
New reactions of the ribosomal RNA precursor of Tetrahymena and the mechanism of self-splicing.
J Mol Biol. 1986 May 5;189(1):143-65 PMID: 2431151

Article Info

Journal

Nucleic acids research

Abbr.

Nucleic Acids Res

ISSN

0305-1048

DOI

10.1093/nar/19.3.605

Published

1991-02-11

Pages

605-9

Language

English

Region

England

NLM ID

0411011

PMCID

PMC333655

Subset

External Links

PubMed source DOI: 10.1093/nar/19.3.605 PMC Full Text View journal details

Analysis Services

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

Contact

WeChat Official Account