The intervening sequence RNA of Tetrahymena is an enzyme.

A shortened form of the self-splicing ribosomal RNA (rRNA) intervening sequence of Tetrahymena thermophila acts as an enzyme in vitro. The enzyme catalyzes the cleavage and rejoining of oligonucleotide substrates in a sequence-dependent manner with Km = 42 microM and kcat = 2 min-1. The reaction mechanism resembles that of rRNA precursor self-splicing. With pentacytidylic acid as the substrate, successive cleavage and rejoining reactions lead to the synthesis of polycytidylic acid. Thus, the RNA molecule can act as an RNA polymerase, differing from the protein enzyme in that it uses an internal rather than an external template. At pH 9, the same RNA enzyme has activity as a sequence-specific ribonuclease.

[1]  T. Cech,et al.  Sites of circularization of the Tetrahymena rRNA IVS are determined by sequence and influenced by position and secondary structure. , 1985, Nucleic acids research.

[2]  T. Cech,et al.  Intermolecular exon ligation of the rRNA precursor of tetrahymena: Oligonucleotides can function as 5′ exons , 1985, Cell.

[3]  T. Cech,et al.  Reactions of the intervening sequence of the Tetrahymena ribosomal ribonucleic acid precursor: pH dependence of cyclization and site-specific hydrolysis. , 1985, Biochemistry.

[4]  M. Belfort,et al.  In vitro expression of the intron-containing gene for T4 phage thymidylate synthase. , 1985, The Journal of biological chemistry.

[5]  T. Cech,et al.  Reversibility of cyclization of the tetrahymena rRNA intervening sequence: implication for the mechanism of splice site choice , 1985, Cell.

[6]  A. Fersht,et al.  Hydrogen bonding in enzymatic catalysis analysed by protein engineering , 1985, Nature.

[7]  N. Pace,et al.  Ribonuclease P catalysis differs from ribosomal RNA self-splicing. , 1985, Science.

[8]  T. Cech,et al.  Coupling of Tetrahymena ribosomal RNA splicing to beta-galactosidase expression in Escherichia coli. , 1985, Science.

[9]  H. Tabak,et al.  Self-splicing of yeast mitochondrial ribosomal and messenger RNA precursors , 1985, Cell.

[10]  T. Cech,et al.  Secondary structure of the circular form of the Tetrahymena rRNA intervening sequence: a technique for RNA structure analysis using chemical probes and reverse transcriptase. , 1985, Proceedings of the National Academy of Sciences of the United States of America.

[11]  A. Lambowitz,et al.  RNA splicing in neurospora mitochondria: Self-splicing of a mitochondrial intron in vitro , 1984, Cell.

[12]  D. Melton,et al.  Efficient in vitro synthesis of biologically active RNA and RNA hybridization probes from plasmids containing a bacteriophage SP6 promoter. , 1984, Nucleic acids research.

[13]  T. Cech,et al.  A labile phosphodiester bond at the ligation junction in a circular intervening sequence RNA. , 1984, Science.

[14]  T. Cech,et al.  Specific interaction between the self-splicing RNA of Tetrahymena and its guanosine substrate: implications for biological catalysis by RNA , 1984, Nature.

[15]  S. Altman,et al.  Catalytic activity of an RNA molecule prepared by transcription in vitro. , 1984, Science.

[16]  N. Pace,et al.  The RNA moiety of ribonuclease P is the catalytic subunit of the enzyme , 1983, Cell.

[17]  R. Waring,et al.  Close relationship between certain nuclear and mitochondrial introns. Implications for the mechanism of RNA splicing. , 1983, Journal of molecular biology.

[18]  M. Zuker,et al.  Secondary structure of the Tetrahymena ribosomal RNA intervening sequence: structural homology with fungal mitochondrial intervening sequences. , 1983, Proceedings of the National Academy of Sciences of the United States of America.

[19]  T. Cech,et al.  Autocatalytic cyclization of an excised intervening sequence RNA is a cleavage–ligation reaction , 1983, Nature.

[20]  R. Waring,et al.  Making ends meet: a model for RNA splicing in fungal mitochondria , 1982, Nature.

[21]  T. Cech,et al.  Self-splicing RNA: Autoexcision and autocyclization of the ribosomal RNA intervening sequence of tetrahymena , 1982, Cell.

[22]  M. Chamberlin,et al.  Bacteriophage SP6-specific RNA polymerase. I. Isolation and characterization of the enzyme. , 1982, The Journal of biological chemistry.

[23]  T. Cech,et al.  In vitro splicing of the ribosomal RNA precursor of tetrahymena: Involvement of a guanosine nucleotide in the excision of the intervening sequence , 1981, Cell.

[24]  P. Modrich,et al.  EcoRI endonuclease. Physical and catalytic properties of the homogenous enzyme. , 1976, The Journal of biological chemistry.

[25]  P. H. Greene,et al.  Restriction and modification of a self-complementary octanucleotide containing the EcoRI substrate. , 1975, Journal of molecular biology.

[26]  C. A. Bunton,et al.  Hydrolysis of bis(2,4-dinitrophenyl) phosphate , 1969 .

[27]  F. Westheimer,et al.  Hydrolysis and Exchange in Esters of Phosphoric Acid1a,b , 1961 .

[28]  F. Long,et al.  Acid Ionization Constants of Alcohols. II. Acidities of Some Substituted Methanols and Related Compounds1,2 , 1960 .

[29]  J. Kumamoto,et al.  Barium Ethylene Phosphate , 1956 .