Click here to close
Hello! We notice that you are using Internet Explorer, which is not supported by Xenbase and may cause the site to display incorrectly.
We suggest using a current version of Chrome,
FireFox, or Safari.
Structural requirements for enzymatic formation of threonylcarbamoyladenosine (t6A) in tRNA: an in vivo study with Xenopus laevis oocytes.
Morin A
,
Auxilien S
,
Senger B
,
Tewari R
,
Grosjean H
.
???displayArticle.abstract???
We have investigated the specificity of the eukaryotic enzymatic machinery that transforms adenosine at position 37 (3' adjacent to anticodon) of several tRNAs into threonylcarbamoyladenosine (t6A37). To this end, 28 variants of yeast initiator tRNAMet and yeast tRNAVal, devoid of modified nucleotide, were produced by in vitro transcription with T7 polymerase of the corresponding synthetic tRNA genes and microinjected into the cytoplasm of Xenopus laevis oocytes. Threonylcarbamoyl incorporation was analyzed in tRNA transcripts mutated in the anticodon loop by substitution, deletion, or Insertion of nucleotides, or in the overall 3D structure of the tRNA by altering critical tertiary interactions. Specifically, we tested the effects of altering ribonucleotides in the anticodon loop, changes of the loop size, perturbations of the overall tRNA 3D structure due to mutations disruptive of the tertiary base pairs, and truncated tRNAs. The results indicate that, in addition to the targeted A37, only U36 was absolutely required. However, A38 in the anticodon loop considerably facilitates the quantitative conversion of A37 into t6A37 catalyzed by the enzymes present in X. laevis. The anticodon positions 34 and 35 were absolutely "neutral" and can accept any of the four canonical nucleotides A, U, C, or G. The anticodon loop size may vary from six to eight nucleotides, and the anticodon stem may have one mismatch pair of the type AxC or GxU at location 30-40 without affecting the efficiency of t6A37 formation and still t6A37 is efficiently formed. Although threonylcarbamoylation of A37 occurred with tRNA having limited perturbations of 3D structure, the overall L-shaped architecture of the tRNA substrate was required for efficient enzymatic conversion of A37 to t6A37. These results favor the idea that unique enzymatic machinery located in the oocyte cytoplasm catalyzes the formation of t6A37 in all U36A37-containing tRNAs (anticodon NNU). Microinjection of the yeast tRNAMeti into the cytoplasm of X. laevis oocytes also revealed the enzymatic activities for several other nucleotide modifications, respectively m1Gg, m2G10, m(2)2G26, m7G46, D47, m5C48/49, and m1A58.
Anderson,
Sequence and organization of the human mitochondrial genome.
1981, Pubmed
Anderson,
Sequence and organization of the human mitochondrial genome.
1981,
Pubmed
Arcari,
The nucleotide sequence of a small (3S) seryl-tRNA (anticodon GCU) from beef heart mitochondria.
1980,
Pubmed
Auxilien,
Mechanism, specificity and general properties of the yeast enzyme catalysing the formation of inosine 34 in the anticodon of transfer RNA.
1996,
Pubmed
Basavappa,
The 3 A crystal structure of yeast initiator tRNA: functional implications in initiator/elongator discrimination.
1991,
Pubmed
Björk,
Genetic dissection of synthesis and function of modified nucleosides in bacterial transfer RNA.
1995,
Pubmed
Bratty,
Characterization of a chemically synthesized RNA having the sequence of the yeast initiator tRNA(Met).
1990,
Pubmed
Chheda,
Biosynthesis of N-(purin-6-ylcarbamoyl)-L-threonine riboside. Incorporation of L-threonine in vivo into modified nucleoside of transfer ribonucleic acid.
1972,
Pubmed
de Bruijn,
A model for the tertiary structure of mammalian mitochondrial transfer RNAs lacking the entire 'dihydrouridine' loop and stem.
1983,
Pubmed
Droogmans,
Enzymatic 2'-O-methylation of the wobble nucleoside of eukaryotic tRNAPhe: specificity depends on structural elements outside the anticodon loop.
1986,
Pubmed
,
Xenbase
Droogmans,
Enzymatic conversion of guanosine 3' adjacent to the anticodon of yeast tRNAPhe to N1-methylguanosine and the wye nucleoside: dependence on the anticodon sequence.
1987,
Pubmed
,
Xenbase
Edqvist,
Enzymatic formation of N2,N2-dimethylguanosine in eukaryotic tRNA: importance of the tRNA architecture.
1995,
Pubmed
,
Xenbase
Elkins,
The enzymatic synthesis of N-(purin-6-ylcarbamoyl)threonine, an anticodon-adjacent base in transfer ribonucleic acid.
1974,
Pubmed
el-Mabrouk,
Very fast identification of RNA motifs in genomic DNA. Application to tRNA search in the yeast genome.
1996,
Pubmed
Grosjean,
Guanosine modifications in runoff transcripts of synthetic transfer RNA-Phe genes microinjected into Xenopus oocytes.
1990,
Pubmed
,
Xenbase
Grosjean,
Enzymatic formation of modified nucleosides in tRNA: dependence on tRNA architecture.
1996,
Pubmed
,
Xenbase
Grosjean,
Structure in tRNA data.
1982,
Pubmed
Grosjean,
The in vivo stability, maturation and aminoacylation of anticodon-substituted Escherichia coli initiator methionine tRNAs.
1987,
Pubmed
,
Xenbase
Grosjean,
Posttranscriptionally modified nucleosides in transfer RNA: their locations and frequencies.
1995,
Pubmed
Harada,
Stable tRNA precursors in HeLa cells.
1984,
Pubmed
Haumont,
Enzymatic formation of queuosine and of glycosyl queuosine in yeast tRNAs microinjected into Xenopus laevis oocytes. The effect of the anticodon loop sequence.
1987,
Pubmed
,
Xenbase
Haumont,
Enzymatic conversion of adenosine to inosine in the wobble position of yeast tRNAAsp: the dependence on the anticodon sequence.
1984,
Pubmed
,
Xenbase
Keith,
Identification and structural characterization of O-beta-ribosyl-(1"----2')-adenosine-5"-phosphate in yeast methionine initiator tRNA.
1990,
Pubmed
Keith,
Mobilities of modified ribonucleotides on two-dimensional cellulose thin-layer chromatography.
1995,
Pubmed
Kiesewetter,
The role of modified purine 64 in initiator/elongator discrimination of tRNA(iMet) from yeast and wheat germ.
1990,
Pubmed
Körner,
N-(purin-6-ylcarbamoyl)threonine: biosynthesis in vitro in transfer RNA by an enzyme purified from Escherichia coli.
1974,
Pubmed
Koski,
Synthesis and maturation of Xenopus laevis methionine tRNA gene transcripts in homologous cell-free extracts.
1982,
Pubmed
,
Xenbase
Kunkel,
Efficient site-directed mutagenesis using uracil-containing DNA.
1991,
Pubmed
Limbach,
Summary: the modified nucleosides of RNA.
1994,
Pubmed
Major,
Reproducing the three-dimensional structure of a tRNA molecule from structural constraints.
1993,
Pubmed
Motorin,
Transfer RNA recognition by the Escherichia coli delta2-isopentenyl-pyrophosphate:tRNA delta2-isopentenyl transferase: dependence on the anticodon arm structure.
1997,
Pubmed
Murgola,
Missense and nonsense suppressors derived from a glycine tRNA by nucleotide insertion and deletion in vivo.
1984,
Pubmed
Peng,
Xenopus laevis: Practical uses in cell and molecular biology. Solutions and protocols.
1991,
Pubmed
,
Xenbase
Perreault,
The synthesis and functional evaluation of RNA and DNA polymers having the sequence of Escherichia coli tRNA(fMet).
1989,
Pubmed
Powers,
Biosynthesis and specific labeling of N-(purin-6-ylcarbamoyl)threonine of Escherichia coli transfer RNA.
1972,
Pubmed
Roberts,
Nucleotide sequence studies of normal and genetically altered glycine transfer ribonucleic acids from Escherichia coli.
1975,
Pubmed
Schweisguth,
On the conformation of the anticodon loops of initiator and elongator methionine tRNAs.
1997,
Pubmed
Senger,
The anticodon triplet is not sufficient to confer methionine acceptance to a transfer RNA.
1992,
Pubmed
Senger,
Yeast tRNA(Met) recognition by methionyl-tRNA synthetase requires determinants from the primary, secondary and tertiary structure: a review.
1996,
Pubmed
Senger,
The presence of a D-stem but not a T-stem is essential for triggering aminoacylation upon anticodon binding in yeast methionine tRNA.
1995,
Pubmed
Simsek,
The primary structure of yeast initiator transfer ribonucleic acid.
1972,
Pubmed
Sprinzl,
Compilation of tRNA sequences and sequences of tRNA genes.
1996,
Pubmed
Steinberg,
A correlation between N2-dimethylguanosine presence and alternate tRNA conformers.
1995,
Pubmed
Steinberg,
Fitting the structurally diverse animal mitochondrial tRNAs(Ser) to common three-dimensional constraints.
1994,
Pubmed
Steinberg,
Structural compensation in atypical mitochondrial tRNAs.
1994,
Pubmed
Szweykowska-Kulinska,
Intron-dependent formation of pseudouridines in the anticodon of Saccharomyces cerevisiae minor tRNA(Ile).
1994,
Pubmed
Tsang,
Sequence specificity of tRNA-modifying enzymes. An analysis of 258 tRNA sequences.
1983,
Pubmed
Watanabe,
Higher-order structure of bovine mitochondrial tRNA(SerUGA): chemical modification and computer modeling.
1994,
Pubmed
Wegnez,
Biochemical research on oogenesis. Nucleotide sequence of initiator tRNA from oocytes and from somatic cells of Xenopus laevis.
1975,
Pubmed
,
Xenbase
Weissenbach,
Effect of threonylcarbamoyl modification (t6A) in yeast tRNA Arg III on codon-anticodon and anticodon-anticodon interactions. A thermodynamic and kinetic evaluation.
1981,
Pubmed
Wolstenholme,
Bizarre tRNAs inferred from DNA sequences of mitochondrial genomes of nematode worms.
1987,
Pubmed
Yarus,
Translational efficiency of transfer RNA's: uses of an extended anticodon.
1982,
Pubmed
Zeevi,
Aminoacylation and nucleoside modification of in vitro synthesised transfer RNA.
1976,
Pubmed
