[6] The use of BD-cellulose in separating transfer RNA's

Gillam, I. C.; Tener, G. M.

doi:10.1016/s0076-6879(71)20008-2

Cited by 45 publications

(12 citation statements)

References 9 publications

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“…Benzoylated DEAE-cellulose [6] was purchased from Boehringer, Japan. [U-14C] serine and [U-14C] leucine were from Amersham, Japan. Oligonucleotide primers used for the polymerase chain reaction (PCR) were synthesized with an automatic DNA synthesizer (Model 391, Applied Biosystems, Japan).…”

Section: Methodsmentioning

confidence: 99%

Characterization of serine and leucine tRNAs in an asporogenic yeastCandida cylindraceaand evolutionary implications of genes for tRNA^SerCAG responsible for translation of a non-universal genetic code

et al. 1994

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Five serine and three leucine isoaceptor tRNAs were purified from the asporogenic yeast Candida cylindracea, in which codon CUG is translated as serine instead of leucine, and their primary structures were determined. From the wobble hypothesis, it was assumed that one of the tRNA(Leu) species (Leu1), with the anticodon CmAA, corresponded to the UUG leucine codon, and that the remaining two leucine tRNAs (Leu2 and Leu3), with the same IAG anticodon sequence would decode the CUU, CUC and CUA codons as leucine, but not the CUG codon; this was clarified by an in vitro translation experiment with C.cylindracea using synthetic mRNAs containing the CUA or CUG codons. One of the serine tRNAs (Ser1) has already been demonstrated to have the anticodon CAG and to be responsible for translation of the codon CUG in C.cylindracea. Three of the other species of tRNA(Ser) (Ser2,3 and 4), with the anticodon sequences cm5UGA, IGA and CGA, can translate all four codons in the UCN codon box, while the remaining species (Ser5), with the anticodon GCU, corresponds to AGU and AGC serine codons. The gene sequences for these five serine and three leucine tRNAs were also determined, with the finding that only tRNA(Ser)CAG (Ser1) has an intron. At least five different types of tRNA(Ser)CAG genes exist in the genome of C.cylindracea. The nucleotide sequences of the flanking regions of these tRNA(Ser)CAG genes indicated that the tRNA(Ser)CAG gene has duplicated at least three times on the genome. The existence of multiple genes for tRNA(Ser)CAG on the genome may account for the observation that codon CUG is used very frequently in C.cylindracea. All of these tRNASerCAG genes contain the CCA sequence in their 3' termini, suggesting the possibility that during their multiplication process in the evolution of the C.cylindracea genome, the tRNA(Ser)CAG molecule was integrated into DNA via reverse transcription.

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Section: Methodsmentioning

confidence: 99%

Characterization of serine and leucine tRNAs in an asporogenic yeastCandida cylindraceaand evolutionary implications of genes for tRNA^SerCAG responsible for translation of a non-universal genetic code

et al. 1994

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“…70-110 Ci/mmol from The Radiochemical Centre, Amersham, UK; uncharged tRNAPhe from Boehringer Mannheim, FRG) was modified by 3-(4'-benzoylphenyl) 3954 propionyl-N-hydroxysuccinimidester as described (Kuechler and Barta, 1977). Non-derivatized tRNA was removed by adsorption of BP-Phe-tRNA to BD-cellulose (Gillam and Tener, 1971). BP-Phe-tRNA was eluted with buffer (10 mM Na-acetate, pH 5, 10 mM MgC92, 1 M NaCI, 30% ethanol).…”

Section: Methodsmentioning

confidence: 99%

Photo-affinity labelling at the peptidyl transferase centre reveals two different positions for the A- and P-sites in domain V of 23S rRNA.

1988

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Photo‐reactive 3‐(4′‐benzoylphenyl)propionyl‐Phe‐tRNA bound to the A‐ or the P‐site was crosslinked to 23S RNA upon irradiation at 320 nm. The sites of reaction were identified as U‐2584 and U‐2585 at the A‐site and A‐2451 and C‐2452 at the P‐site. Minor crosslinks from both sites were observed at nucleotides A‐2503 to U‐2506. All sites identified lie in close proximity according to the secondary structure model and constitute part of the highly conserved loop region of domain V. Antibiotics known to inhibit peptidyl transferase activity had a pronounced effect on photo‐crosslinking. In addition, tetracycline was also shown to photo‐crosslink to this region. These experiments permit a dissection of the peptidyl transferase region on the 23S RNA into two distinct areas for the A‐ and P‐site.

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“…Crude tRNA obtained from the liver of inbred Buffalo rats (7) was fractionated on a BD-cellulose column (8). The first major glutamine accepting peak (hereafter designated as tRNAG(ln) was further chromatographed on Sepharose 4B (9), followed by repeated polyacrylamide gel electrophoresis (10).…”

Section: Methodsmentioning

confidence: 99%

The nucleotide sequence of a major glutamine tRNA from rat liver

et al. 1983

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[6] The use of BD-cellulose in separating transfer RNA's

Cited by 45 publications

References 9 publications

Characterization of serine and leucine tRNAs in an asporogenic yeastCandida cylindraceaand evolutionary implications of genes for tRNA^SerCAG responsible for translation of a non-universal genetic code

Characterization of serine and leucine tRNAs in an asporogenic yeastCandida cylindraceaand evolutionary implications of genes for tRNA^SerCAG responsible for translation of a non-universal genetic code

Photo-affinity labelling at the peptidyl transferase centre reveals two different positions for the A- and P-sites in domain V of 23S rRNA.

The nucleotide sequence of a major glutamine tRNA from rat liver

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[6] The use of BD-cellulose in separating transfer RNA's

Cited by 45 publications

References 9 publications

Characterization of serine and leucine tRNAs in an asporogenic yeastCandida cylindraceaand evolutionary implications of genes for tRNASerCAG responsible for translation of a non-universal genetic code

Characterization of serine and leucine tRNAs in an asporogenic yeastCandida cylindraceaand evolutionary implications of genes for tRNASerCAG responsible for translation of a non-universal genetic code

Photo-affinity labelling at the peptidyl transferase centre reveals two different positions for the A- and P-sites in domain V of 23S rRNA.

The nucleotide sequence of a major glutamine tRNA from rat liver

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Characterization of serine and leucine tRNAs in an asporogenic yeastCandida cylindraceaand evolutionary implications of genes for tRNA^SerCAG responsible for translation of a non-universal genetic code

Characterization of serine and leucine tRNAs in an asporogenic yeastCandida cylindraceaand evolutionary implications of genes for tRNA^SerCAG responsible for translation of a non-universal genetic code