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.
A cell-free plant extract for accurate pre-tRNA processing, splicing and modification.
Stange N
,
Beier H
.
???displayArticle.abstract???
An intron-containing tobacco tRNA(Tyr) precursor synthesized in a HeLa cell nuclear extract has been used to develop a cell-free processing and splicing system from wheat germ. Removal of 5' and 3' flanking sequences, accurate excision of the intervening sequence, ligation of the resulting tRNA halves, addition of the 3'-terminal CCA sequence and modification of seven nucleosides were achieved in appropriate wheat germ S23 and S100 extracts. The maturation of pre-tRNA(Tyr) in these extracts resembles the pathway observed in vivo for tRNA biosynthesis in Xenopus oocytes and yeast in that processing of the flanks precedes intron excision. Most of the modified nucleosides (m2(2) G, psi 35, psi 55, m7G and m1A) are introduced into the intron-containing pre-tRNA with mature ends, whereas two others (m1G and psi 39) are only found in the mature tRNA(Tyr). Processing and splicing proceed very efficiently in the wheat germ extracts, leading to complete maturation of 5' and 3' ends followed by about 65% conversion to mature tRNA(Tyr) under our standard conditions. The activity of the wheat germ endonuclease is stimulated 3-fold by the non-ionic detergent Triton X-100. All previous attempts to demonstrate the presence of a splicing endonuclease in wheat germ had failed (Gegenheimer et al., 1983). Hence, this is the first cell-free plant extract which supports pre-tRNA processing and splicing in vitro.
Beier,
The molecular basis for the differential translation of TMV RNA in tobacco protoplasts and wheat germ extracts.
1984, Pubmed
Beier,
The molecular basis for the differential translation of TMV RNA in tobacco protoplasts and wheat germ extracts.
1984,
Pubmed
Beier,
UAG readthrough during TMV RNA translation: isolation and sequence of two tRNAs with suppressor activity from tobacco plants.
1984,
Pubmed
Bradford,
A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding.
1976,
Pubmed
Brown,
Accurate in vitro splicing of two pre-mRNA plant introns in a HeLa cell nuclear extract.
1986,
Pubmed
Brown,
A catalogue of splice junction and putative branch point sequences from plant introns.
1986,
Pubmed
Cheng,
Fractionation and characterization of a yeast mRNA splicing extract.
1986,
Pubmed
Colby,
Yeast tRNA precursor mutated at a splice junction is correctly processed in vivo.
1981,
Pubmed
Dignam,
Accurate transcription initiation by RNA polymerase II in a soluble extract from isolated mammalian nuclei.
1983,
Pubmed
Domdey,
Studies on the primary and secondary structure of potato spindle tuber viroid: products of digestion with ribonuclease A and ribonuclease T1, and modification with bisulfite.
1978,
Pubmed
Engelke,
Nucleolytic processing of a tRNAArg-tRNAAsp dimeric precursor by a homologous component from Saccharomyces cerevisiae.
1985,
Pubmed
Etcheverry,
A precursor to a minor species of yeast tRNASer contains an intervening sequence.
1979,
Pubmed
Filipowicz,
Origin of splice junction phosphate in tRNAs processed by HeLa cell extract.
1983,
Pubmed
,
Xenbase
Gegenheimer,
An RNA ligase from wheat germ which participates in transfer RNA splicing in vitro.
1983,
Pubmed
Green,
Purification and properties of a mammalian tRNA pseudouridine synthase.
1982,
Pubmed
Greer,
Mechanism of action of a yeast RNA ligase in tRNA splicing.
1983,
Pubmed
Greer,
Assembly of a tRNA splicing complex: evidence for concerted excision and joining steps in splicing in vitro.
1986,
Pubmed
Gross,
Nucleotide sequence and secondary structure of citrus exocortis and chrysanthemum stunt viroid.
1982,
Pubmed
Hartmuth,
In vitro processing of a plant pre-mRNA in a HeLa cell nuclear extract.
1986,
Pubmed
Hopper,
tRNA synthesis: identification of in vivo precursor tRNAs from parental and mutant yeast strains.
1981,
Pubmed
,
Xenbase
Johnson,
The yeast tRNATyr gene intron is essential for correct modification of its tRNA product.
1983,
Pubmed
Knapp,
Transcription and processing of intervening sequences in yeast tRNA genes.
1978,
Pubmed
Konarska,
Formation of a 2'-phosphomonoester, 3',5'-phosphodiester linkage by a novel RNA ligase in wheat germ.
,
Pubmed
Koski,
Synthesis and maturation of Xenopus laevis methionine tRNA gene transcripts in homologous cell-free extracts.
1982,
Pubmed
,
Xenbase
Koski,
An in vitro RNA polymerase III system from S. cerevisiae: effects of deletions and point mutations upon SUP4 gene transcription.
1982,
Pubmed
,
Xenbase
Laski,
Characterization of tRNA precursor splicing in mammalian extracts.
1983,
Pubmed
,
Xenbase
Laski,
Expression of a X. laevis tRNATyr gene in mammalian cells.
1982,
Pubmed
,
Xenbase
Melton,
Order and intracellular location of the events involved in the maturation of a spliced tRNA.
1980,
Pubmed
,
Xenbase
Nishikura,
RNA processing in microinjected Xenopus oocytes. Sequential addition of base modifications in the spliced transfer RNA.
1981,
Pubmed
,
Xenbase
O'Farrell,
Structure and processing of yeast precursor tRNAs containing intervening sequences.
1978,
Pubmed
Ogden,
In vitro transcription and processing of a yeast tRNA gene containing an intervening sequence.
1979,
Pubmed
,
Xenbase
Peebles,
Precise excision of intervening sequences from precursor tRNAs by a membrane-associated yeast endonuclease.
1983,
Pubmed
Peebles,
Splicing of yeast tRNA precursors: a two-stage reaction.
1979,
Pubmed
Rooney,
Processing of mammalian tRNA transcripts in vitro: different pre-tRNAs are processed along alternative pathways that contain a common rate-limiting step.
1986,
Pubmed
Schaefer,
Nucleotide modification in vitro of the precursor of transfer RNA of Escherichia coli.
1973,
Pubmed
Schwartz,
Enzymatic mechanism of an RNA ligase from wheat germ.
1983,
Pubmed
Silberklang,
Use of in vitro 32P labeling in the sequence analysis of nonradioactive tRNAs.
1979,
Pubmed
Sprinzl,
Compilation of tRNA sequences and sequences of tRNA genes.
1987,
Pubmed
Standring,
Yeast tRNA3Leu gene transcribed and spliced in a HeLa cell extract.
1981,
Pubmed
,
Xenbase
Stange,
A gene for the major cytoplasmic tRNATyr from Nicotiana rustica contains a 13 nucleotides long intron.
1986,
Pubmed
Strobel,
Effect of intron mutations on processing and function of Saccharomyces cerevisiae SUP53 tRNA in vitro and in vivo.
1986,
Pubmed
Tyc,
Ligation of endogenous tRNA 3' half molecules to their corresponding 5' halves via 2'-phosphomonoester,3',5'-phosphodiester bonds in extracts of Chlamydomonas.
1983,
Pubmed
van Tol,
A human and a plant intron-containing tRNATyr gene are both transcribed in a HeLa cell extract but spliced along different pathways.
1987,
Pubmed
,
Xenbase
Walden,
Purification and properties of guanine, queuine-tRNA transglycosylase from wheat germ.
1982,
Pubmed
Waldron,
Plant tRNA genes: putative soybean genes for tRNAasp and tRNAmet.
1985,
Pubmed
