Warning: Undefined array key "mm" in /www/wwwroot/www.ai-bt.com/si.php on line 10 Deprecated: trim(): Passing null to parameter #1 ($string) of type string is deprecated in /www/wwwroot/www.ai-bt.com/si.php on line 10 Biochemical and physical characterization of an unmodified yeast phenylalanine transfer RNA transcribed in vitro.

Literature DB >> 3277187

Biochemical and physical characterization of an unmodified yeast phenylalanine transfer RNA transcribed in vitro.

Abstract

A recombinant plasmid was constructed with six synthetic DNA oligomers such that the DNA sequence corresponding to yeast tRNA(Phe) is flanked by a T7 promoter and a BstNI restriction site. Runoff transcription of the BstNI-digested plasmid with T7 RNA polymerase gives an unmodified tRNA of the expected sequence having correct 5' and 3' termini. This tRNA(Phe) transcript can be specifically aminoacylated by yeast phenylalanyl-tRNA synthetase and has a Km only 4-fold higher than that of the native yeast tRNA(Phe). The Km is independent of Mg2+ concentration, whereas the Vmax is very dependent on Mg2+ concentration. Comparison of the melting profiles of the native and the unmodified tRNA(Phe) at different Mg2+ concentrations suggests that the unmodified tRNA(Phe) has a less stable tertiary structure. Using one additional DNA oligomer, a mutant plasmid was constructed having a guanosine to thymidine change at position 20 in the tRNA gene. A decrease in Vmax/Km by a factor of 14 for aminoacylation of the mutant tRNA(Phe) transcript is observed.

Entities: Chemical Species

Mesh：

Substances：

Year: 1988 PMID： 3277187 PMCID： PMC279695 DOI： 10.1073/pnas.85.4.1033

Source DB: PubMed Journal: Proc Natl Acad Sci U S A ISSN： 0027-8424 Impact factor: 11.205

26 in total

1. Aminoacylation and nucleoside modification of in vitro synthesised transfer RNA.

Authors: M Zeevi; V Daniel
Journal: Nature Date: 1976-03-04 Impact factor: 49.962

2. Structural analysis of spermine and magnesium ion binding to yeast phenylalanine transfer RNA.

Authors: G J Quigley; M M Teeter; A Rich
Journal: Proc Natl Acad Sci U S A Date: 1978-01 Impact factor: 11.205

3. Function of N2 methylguanine in phenylalanine transfer RNA.

Authors: B Roe; M Michael; B Dudock
Journal: Nat New Biol Date: 1973-12-05

4. Kethoxal inactivation of three transfer ribonucleic acids chargeable by yeast phenylalanyl transfer ribonucleic acid synthetase.

Authors: M Litt; C M Greenspan
Journal: Biochemistry Date: 1972-04-11 Impact factor: 3.162

5. Aminoacylation of anticodon loop substituted yeast tyrosine transfer RNA.

Authors: L Bare; O C Uhlenbeck
Journal: Biochemistry Date: 1985-04-23 Impact factor: 3.162

6. In vitro construction of yeast tRNAAsp variants: nucleotide substitutions and additions in T-stem and T-loop.

Authors: P Carbon; J P Ebel
Journal: Nucleic Acids Res Date: 1987-03-11 Impact factor: 16.971

7. Structural domains of transfer RNA molecules.

Authors: G J Quigley; A Rich
Journal: Science Date: 1976-11-19 Impact factor: 47.728

8. Oligoribonucleotide synthesis using T7 RNA polymerase and synthetic DNA templates.

Authors: J F Milligan; D R Groebe; G W Witherell; O C Uhlenbeck
Journal: Nucleic Acids Res Date: 1987-11-11 Impact factor: 16.971

9. RNA-ligant interactions. (I) Magnesium binding sites in yeast tRNAPhe.

Authors: S R Holbrook; J L Sussman; R W Warrant; G M Church; S H Kim
Journal: Nucleic Acids Res Date: 1977-08 Impact factor: 16.971

10. Functional mutants of phenylalanine transfer RNA from Escherichia coli.

Authors: J Vacher; M Springer; R H Buckingham
Journal: EMBO J Date: 1985-02 Impact factor: 11.598

267 in total

1. Identity elements in bovine tRNA(Trp) required for the specific stimulation of gelonin, a plant ribosome-inactivating protein.

Authors: M Brigotti; D Carnicelli; A Pallanca; S Rizzi; P Accorsi; L Montanaro; S Sperti
Journal: RNA Date: 1999-10 Impact factor: 4.942

2. The RNase P associated with HeLa cell mitochondria contains an essential RNA component identical in sequence to that of the nuclear RNase P.

Authors: R S Puranam; G Attardi
Journal: Mol Cell Biol Date: 2001-01 Impact factor: 4.272

10. The catalytic flexibility of tRNAIle-lysidine synthetase can generate alternative tRNA substrates for isoleucyl-tRNA synthetase.

Authors: Scott P Salowe; Judyann Wiltsie; Julio C Hawkins; Lisa M Sonatore
Journal: J Biol Chem Date: 2009-02-19 Impact factor: 5.157

Biochemical and physical characterization of an unmodified yeast phenylalanine transfer RNA transcribed in vitro.

1. Aminoacylation and nucleoside modification of in vitro synthesised transfer RNA.

2. Structural analysis of spermine and magnesium ion binding to yeast phenylalanine transfer RNA.

3. Function of N2 methylguanine in phenylalanine transfer RNA.

4. Kethoxal inactivation of three transfer ribonucleic acids chargeable by yeast phenylalanyl transfer ribonucleic acid synthetase.

5. Aminoacylation of anticodon loop substituted yeast tyrosine transfer RNA.

6. In vitro construction of yeast tRNAAsp variants: nucleotide substitutions and additions in T-stem and T-loop.

7. Structural domains of transfer RNA molecules.

8. Oligoribonucleotide synthesis using T7 RNA polymerase and synthetic DNA templates.

9. RNA-ligant interactions. (I) Magnesium binding sites in yeast tRNAPhe.

10. Functional mutants of phenylalanine transfer RNA from Escherichia coli.

1. Identity elements in bovine tRNA(Trp) required for the specific stimulation of gelonin, a plant ribosome-inactivating protein.

2. The RNase P associated with HeLa cell mitochondria contains an essential RNA component identical in sequence to that of the nuclear RNase P.

3. An engineered class I transfer RNA with a class II tertiary fold.

4. Binding and cross-linking of tmRNA to ribosomal protein S1, on and off the Escherichia coli ribosome.

5. Ala-His mediated peptide bond formation revisited.

6. Preparation and activity of synthetic unmodified mammalian tRNAi(Met) in initiation of translation in vitro.

7. Escherichia coli DbpA is an RNA helicase that requires hairpin 92 of 23S rRNA.

8. A one-step method for in vitro production of tRNA transcripts.

9. Identification and characterization of a tRNA decoding the rare AUA codon in Haloarcula marismortui.

10. The catalytic flexibility of tRNAIle-lysidine synthetase can generate alternative tRNA substrates for isoleucyl-tRNA synthetase.