Selective removal of protecting groups for phosphomonoesters of nucleotides by anodic oxidation.

Ohtsuka, Eiko; Miyake, Toru; Ikehara, Morio; Matsumoto, A.; Ohmori, Hidenobu

doi:10.1248/cpb.27.2242

Cited by 5 publications

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“…We began by synthesizing terminal fragments of the tRNA (11)(12)(13)(14)(15)(16) and examined the ability of RNA ligase (17)(18)(19) to join these fragments. The 5'-terminal icosanucleotide (20), the tetradecanucleotide (bases 21-34) (21), and the 3'-heptadecanucleotide (22) have been obtained by this method.…”

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confidence: 99%

Total synthesis of a RNA molecule with sequence identical to that of Escherichia coli formylmethionine tRNA.

Ohtsuka¹,

Tanaka²,

Tanaka³

et al. 1981

Proc. Natl. Acad. Sci. U.S.A.

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A RNA molecule has been synthesized that is identical in sequence to Escherichia coli tRNA; except that it lacks the base modifications present in the E. coli tRNA. This was achieved by enzymatic joining of chemically synthesized oligonucleotides with chain lengths.of3-10 which were synthesized by the phosphodiester or phosphotriester method. First, quarter molecules of tRNA were constructed by joining of chemically synthesized fragments with RNA ligase. The 5'-quarter molecule (bases 1-20) served as an acceptor in joining reactions with the 3',5'-bisphosphorylated donor molecule (bases 21-34). The 5'-half molecule thus obtained was treated with phosphatase andjoined to the 3'-half molecule which was prepared by ligation of the other quarter molecules (bases 35-60, acceptor; bases 61-77, donor) followed by 5'-phosphorylation with polynucleotide kinase. The synthetic tRNA was characterized by oligonucleotide pattern and was partially active in aminoacylation with E. coli methionyl-tRNA synthetase.Chemical synthesis ofnucleic acids has been a challenging problem in organic chemistry since the structure ofthe nucleic acids was elucidated. Chemical methods to synthesize short ribo-and deoxyribopolynucleotides with defined sequences were established in early 1960s, and those oligonucleotides were important in the elucidation of the genetic code (1). Discovery of DNA ligase allowed the synthesis of bihelical DNAs from chemically synthesized deoxyribopolynucleotides. With this chemical-enzymatic method the genes for yeast alanine tRNA (2) and Escherichia coli tyrosine tRNA precursor (3) have been synthesized; the latter was the first synthetic functional DNA molecule. Genes for peptides have also been synthesized by the same approach, and the methods for joining double-stranded DNA pieces with protruding ends have been used in various recently developed reactions for genetic manipulations.Although tRNAs are the smallest nucleic acids with unique functions, their synthesis has been difficult until recently, mainly because of the lack ofgood synthetic methods for larger .oligoribonucleotides as well as a lack ofjoining enzymes. After the primary structure of yeast alanine tRNA had been determined (4), the nona-and hexanucleotide corresponding to the terminal sequence of this tRNA were synthesized by phosphodiester block condensation. These fragments in turn were used to form reconstituted molecules with natural tRNA fragments derived by RNase digestions. However, aminoacylation was not possible because the synthetic fragments were too small to form sufficiently stable complexes for recognition by the alanyl-tRNA synthetase (5). The discovery of RNA ligase (6) and its ability to join single-stranded oligoribonucleotides (7) made it possible to elongate synthetic RNA fragments to yield larger molecules such as tRNAs.The initiator methionine tRNA of prokaryotes has a special role in protein biosynthesis, which. manifests itself in several unique properties of that tRNA (8). It was also the subject of detailed modif...

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confidence: 99%

Total synthesis of a RNA molecule with sequence identical to that of Escherichia coli formylmethionine tRNA.

Ohtsuka¹,

Tanaka²,

Tanaka³

et al. 1981

Proc. Natl. Acad. Sci. U.S.A.

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“…The reaction mixture was added to 5% potassium acetate (15 ml) with cooling and stirred at room temperature for 1 hr. The product (14) was extracted with chloroform (10 ml) 3 portions, washed with water (10 ml) 3 portions, concentrated with pyridine and with toluene.…”

Section: L Nucleic Acids Researchmentioning

confidence: 99%

“…The mixture was kept at room temperature overnight, decanted and the aqueous phase was filtered to remove dicyclohexylurea, DCC was removed with n-hexane (30 ml). The nucleotide (16) was extracted with chloroform (15 ml) 3 portions, washed with 0.1M triethylammonium bicarbonate (pH 7.5) (20 ml) 3 portions, concentrated with pyridine ahid precipitated with n-pentane (100 ml) from its solution in chloroform (5 ml). The yield was 99% (17400 A261, 0.79 mmol).…”

Section: L Nucleic Acids Researchmentioning

confidence: 99%

“…The phosphotriester approach using 2'-O-(o-nitrobenzylnucleoside)3'-phosphorodianilidates was also employed in the synthesis of a eukaryotic initiator tRNA loop IV heptanucleotide.12 In the present paper we report a synthesis of 5'pentanucleotides from E.coli tRNAfMet, C-G-C-G-Gp (bases [1][2][3][4][5], U-G-C-G-Gp (base 1 transition analog), pG-G-C-G-Gp (base 1 transversion analog) and pG-G-s4U-G-Gp (bases 6-10) using 2'-O-(onitrobenzyl nucleoside)-3'-p-chlorophenylphosphoranilidates.10' 13 Protected pGp was prepared for insertion of the 5'-phosphomonoester in the synthesis of the pentanucleotide, pG-G-C-G-Gp and the pentanucleotide was converted to the s4U containing oligonucleotide by treatment with liquid hydrogen sulfide.15 Synthesis of C-G-C-G-Gp (10) and U-G-C-G-Gp (11) For the synthesis of pentanucleotides (8,9), a common intermediate tetranucleotide (5) was prepared by condensation of two a InfnrM Retival Umed I Falconb.rg Court London W1VSFG England Nucleic Acids Research…”

Section: Introductionmentioning

confidence: 99%

“…The reaction conditions for synthesis of protected oligonucleotides are summarized in Table I. Protected GpCp (3) and GpGp (4) were obtained by condensation of mononucleotides as described previously.°The tetranucleotide (5) was condensed with two equivalents of (6) using TPSNI to give protected C-G-C-G-Gp (8). The product was isolated by chromatography on silica gel.…”

Section: Introductionmentioning

confidence: 99%

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Studies on transfer ribonucleic acids and related compounds. XXXII. Synthesis of ribonucleotides corresponding to residues 1-5 and 6-10 of tRNA^fMetfrom E.coli and their base conversion analogs

Ohtsuka¹,

Tanaka²,

Ikehara³

1979

Nucl Acids Res

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E. Coli tRNAfMet fragments, C-G-C-G-Gp (bases 1-5), U-G-C-G-Gp (base 1 transition, analog) pG-G-C-G-Gp (base 1 transversion analog) and pG-G-s4U-G-Gp (bases 6-10) were synthesized by triester methods using 2'-O-(o-nitrobenzyl) nucleotides including a 3',5'-bisphosphorylated guanosine derivative. The s4U containing pentanucleotide was derived from the pG-G-C-G-Gp by treatment with liquid hydrogen sulfide.

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Protection for the Phosphate Group

Wuts

Greene²

2006

Greene's Protective Groups in Organic Synthesis

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Selective removal of protecting groups for phosphomonoesters of nucleotides by anodic oxidation.

Cited by 5 publications

References 0 publications

Total synthesis of a RNA molecule with sequence identical to that of Escherichia coli formylmethionine tRNA.

Total synthesis of a RNA molecule with sequence identical to that of Escherichia coli formylmethionine tRNA.

Studies on transfer ribonucleic acids and related compounds. XXXII. Synthesis of ribonucleotides corresponding to residues 1-5 and 6-10 of tRNA^fMetfrom E.coli and their base conversion analogs

Protection for the Phosphate Group

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Selective removal of protecting groups for phosphomonoesters of nucleotides by anodic oxidation.

Cited by 5 publications

References 0 publications

Total synthesis of a RNA molecule with sequence identical to that of Escherichia coli formylmethionine tRNA.

Total synthesis of a RNA molecule with sequence identical to that of Escherichia coli formylmethionine tRNA.

Studies on transfer ribonucleic acids and related compounds. XXXII. Synthesis of ribonucleotides corresponding to residues 1-5 and 6-10 of tRNAfMetfrom E.coli and their base conversion analogs

Protection for the Phosphate Group

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Studies on transfer ribonucleic acids and related compounds. XXXII. Synthesis of ribonucleotides corresponding to residues 1-5 and 6-10 of tRNA^fMetfrom E.coli and their base conversion analogs