Oligoribonucleotide synthesis. X. An improved synthesis of the anticodon loop region of methionine transfer ribonucleic acid from <i>E</i>. <i>coli</i>

Werstiuk, Eva S.; Neilson, Thomas

doi:10.1139/v76-381

Cited by 37 publications

(12 citation statements)

References 8 publications

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“…The existing phosphotriester synthesis [12,13] could already provide all acceptor oligomers (between four and six residues). With the extension described in this paper, synthetic oligoribonucleotide 3'-monophosphates can be obtained in good yield.…”

Section: Discussionmentioning

confidence: 99%

“…Presently, only chemical synthesis of oligoribonucleotides provides the versatility, both in scale of preparation and in variety of sequences potentially available, for the systematic investigation of any facet of the mechanism of protein synthesis. The existing phosphotriester synthesis [12,13] could already provide all acceptor oligomers (between four and six residues). With the extension described in this paper, synthetic oligoribonucleotide 3'-monophosphates can be obtained in good yield.…”

Section: Discussionmentioning

confidence: 99%

“…All oligoribonucleotides used in this study were prepared by the general phosphotriester synthesis developed by Neilson and his associates [12,13]. At the present time, only this synthesis provides satisfactory coupling to guanosine residues and so can supply sufficient quantities of oligomers of defined sequence beyond the trimer level.…”

Section: Chemical Synthesis Of Oligoribonucleotidesmentioning

confidence: 99%

“…Protected oligomers were completely deblocked using the three-step procedure described previously [13]. Yields for the 3'-monophosphates were markedly lower than those for their parents, possibly due to increased adsorption on the Chelex 100 resin.…”

Section: Characterization Of Oligoribonucleotidesmentioning

confidence: 99%

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Synthesis of Biologically Active Portions of an Intercistronic Region by Use of a New 3′‐Phosphate Incorporation Method to Protect 3′‐OH and Their Binding to Ribosomes

Neilson

Gregoire

Fraser

et al. 1979

European Journal of Biochemistry

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To examine the influence of bases contiguous to a starter codon, a rapid means of assembling biologically active regions corresponding to portions, or to analogues of portions of intercistronic regions is desirable. To do this, a chemical method for the specific insertion of 3′‐monophosphate groups on to chemically synthesized trinucleotides, tetranucleotides and hexanucleotides of defined sequence bearing different bases at the 3′ terminus has been devised. The method involved phosphorylation of blocked oligoribonucleotides synthesized by a phosphotriester method. The deblocked oligoribonucleotides were phosphorylated at the 5′ end with T4‐induced polynucleotide kinase. The products of this kinase reaction served as“donors” in RNA ligase reactions. The [32P]pC‐A‐U‐A‐U‐Gp, [32P]pA‐U‐Gp, [32P]pU‐A‐A, [32P]pA‐G‐G‐Ap, [32P]pC‐U‐U‐Ap and [32P]pU‐C‐C‐Up “donors” were used to synthesize A‐G‐G‐A[32P]pC‐A‐U‐A‐U‐Gp, U‐C‐C‐U[32P]pC‐A‐U‐A‐U‐Gp, U‐A‐A[32P]pA‐U‐G, A‐U‐G[32P]pU‐A‐A, U‐A‐A‐G[32P]pA‐G‐G‐Ap, U‐C‐C‐U[32P]pC‐U‐U‐Ap and A‐U‐U‐C[32P]pU‐C‐C‐Up indicating that the method functions with all bases. A‐U‐Gp, PA‐U‐Gp and pC‐A‐U‐A‐U‐Gp were isolated free of reactants and, along with PA‐U‐G, were all shown to promote the formation of translational initiation complexes. A‐G‐G‐A[32P]pC‐A‐U‐A‐U‐Gp, which corresponds to the 5′‐terminal portion of the intracistronic region of the maturation protein of bacteriophage Qβ, bound more efficiently to Escherichia coli ribosomes than the U‐C‐C‐U[32P]pC‐A‐U‐A‐U‐Gp, [32P]pC‐A‐U‐A‐U‐Gp, [32P]pA‐U‐G‐U‐A‐A or [32P]pU‐A‐A‐A‐U‐G controls. The results suggested that the content and number of residues at the 5′ terminus attached to A‐U‐G affect binding of oligonucleotides to ribosomes; purine nucleosides appear to be more effective than pyrimidine nucleosides in this regard.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Chemical Synthesis Of Oligoribonucleotidesmentioning

confidence: 99%

Section: Characterization Of Oligoribonucleotidesmentioning

confidence: 99%

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Synthesis of Biologically Active Portions of an Intercistronic Region by Use of a New 3′‐Phosphate Incorporation Method to Protect 3′‐OH and Their Binding to Ribosomes

Neilson

Gregoire

Fraser

et al. 1979

European Journal of Biochemistry

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“…The oligoribonucleotides used in this study wcrc chemically synthesized by a phosphotricster method (16)(17)(18).…”

Section: Methodsmentioning

confidence: 99%

The application of double-quantum and spin echo nuclear magnetic resonance spectroscopy to the assignment of ¹H chemical shifts of RNA oligomers

Hughes¹,

Neilson²,

Coddington³

et al. 1984

Can. J. Chem.

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, 1214 (1984).Specific assignment of the non-exchangeable base and ribosc anomcric protons is difficult for oligoribonucleotides with a high pyrimidine content. Problems arise from the narrow range of pyrimidine H-6 chemical shifts and thc ovcrlap of pyrimidine H-5 rcsonanccs with thc ribose anomeric proton signals. To solve thcsc problems two multiple pulsc methods werc used which depend on the scalar coupling betwccn H-6 and H-5. The first method was double-quantum nmr which established the connectivity bctwecn specific pyrimidine H-6 and H-5 rcsonances. A rcfocussing pulse was added to the end of the doublequantum pulse sequence to improve the identification of several H-6 and H-5 connectivities in a single expcriment. The second procedure involved hornonuclcar decouplcd spin ccho nmr to remove H-5 signals sclcctivcly from the ribose anomeric proton region. These techniques are illustrated by the assignment of the spectra of the oligoribonucleotides UpUpC, UpUpU, GpCpUpC, and UpApGpCpUpU.

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Nearest‐neighbor and next‐nearest‐neighbor effects in the proton NMR spectra of the oligoribonucleotides ApGpX and CpApX

Everett¹,

Hughes²,

Bell³

et al. 1980

Biopolymers

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SynopsisThe variable-temperature proton nmr spectra of the oligoribonucleotides in the series CpApX and the series ApGpX, X = A, G, C, U, together with the parent dimers CpA and ApG have been measured. A complete analysis of all the nonexchangeable base proton resonances and ribose H-1' proton resonances was made. The presence of trends in the shielding abilities of the various bases at both the nearest-neighbor and next-nearest-neighbor positions were identified. The observed shieldings could be used to predict the chemical shifts of protons in related systems. Based on the empirical results from ribodinucleoside monophosphates, the temperature-dependent behavior of the J1.2, coupling constants of the triribonucleotides suggested that the compounds in the CpApX series stacked from the 5'-end to the 3'-end, while those in the ApGpX series stacked from the 3'-end to the 5'-end.

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Oligoribonucleotide synthesis. X. An improved synthesis of the anticodon loop region of methionine transfer ribonucleic acid from E. coli

Cited by 37 publications

References 8 publications

Synthesis of Biologically Active Portions of an Intercistronic Region by Use of a New 3′‐Phosphate Incorporation Method to Protect 3′‐OH and Their Binding to Ribosomes

Synthesis of Biologically Active Portions of an Intercistronic Region by Use of a New 3′‐Phosphate Incorporation Method to Protect 3′‐OH and Their Binding to Ribosomes

The application of double-quantum and spin echo nuclear magnetic resonance spectroscopy to the assignment of ¹H chemical shifts of RNA oligomers

Nearest‐neighbor and next‐nearest‐neighbor effects in the proton NMR spectra of the oligoribonucleotides ApGpX and CpApX

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Oligoribonucleotide synthesis. X. An improved synthesis of the anticodon loop region of methionine transfer ribonucleic acid from E. coli

Cited by 37 publications

References 8 publications

Synthesis of Biologically Active Portions of an Intercistronic Region by Use of a New 3′‐Phosphate Incorporation Method to Protect 3′‐OH and Their Binding to Ribosomes

Synthesis of Biologically Active Portions of an Intercistronic Region by Use of a New 3′‐Phosphate Incorporation Method to Protect 3′‐OH and Their Binding to Ribosomes

The application of double-quantum and spin echo nuclear magnetic resonance spectroscopy to the assignment of 1H chemical shifts of RNA oligomers

Nearest‐neighbor and next‐nearest‐neighbor effects in the proton NMR spectra of the oligoribonucleotides ApGpX and CpApX

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The application of double-quantum and spin echo nuclear magnetic resonance spectroscopy to the assignment of ¹H chemical shifts of RNA oligomers