An analytical method for the separation of sugar-methylated ribonucleosides from base-methylated and nonmethylated ribonucleosides

Al-Arif, Adhid; Sporn, Michael B.

doi:10.1016/0003-2697(72)90091-7

Cited by 27 publications

(10 citation statements)

References 15 publications

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“…In a typical experiment, a known aliquot of the enzymatic hydrolysate was chromatographed essentially according to the procedure by Arif & Sporn (1972), except that the Whatman paper No.1 was not pre-impregnated with ammonium borate. After a 14 hrs development at 22 °C, the chromatogram was removed from the chamber, air-dried, and the resolved nucleosides were identified as above.…”

Section: Methodsmentioning

confidence: 99%

5-methylcytosine content in the vertebrate deoxyribonucleic acids: Species specificity

Kothari

Shankar

1976

J Mol Evol

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RNA-free native DNA samples, isolated by four methods, from different vertebrate tissues and species, were hydrolyzed chemically and enzymatically and analyzed by paper chromatography to estimate the base composition. It was noted that (i) all the DNA preparations analyzed contained 5-methylcytosine, (ii) on the basis of mole percent of 5-methylcytosine, the composition of DNA varied in different species, but not so much in different tissues of the same species, (iii) the method of DNA hydrolysis, but not the method of deproteinization, affected the mole percent of 5-methylcytosine, and (iv) no 5-hydroxymethylcytosine (5-HMC) was detected in any of the DNA preparations analyzed.

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Section: Methodsmentioning

confidence: 99%

5-methylcytosine content in the vertebrate deoxyribonucleic acids: Species specificity

Kothari

Shankar

1976

J Mol Evol

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“…N o attempt was made to resolve 2'-0alkyl and 3'-O-alkyl nucleosides. Solvent I is 1 -butanol-ethanol-water (80:10:25) on Whatman 3 MM, descending 16-24 h. Solvent I1 is 1 -butanol-concentrated ammonium hydroxide-0.8 M boric acid (200:0.8:27) on Whatman 3 MM, saturated with 0.1 M ammonium borate and dried, descending 22 h (AI- Arif and Sporn, 1972). Solvent 111 is l-butanol-concentrated ammonium hydroxide-H20 (85:2:12) on Whatman 3 MM, descending 18 h. Solvent IV is acetone-benzene (2: 1) on activated silica gel sheets, ascending 2-3 h (KuSmierek and Singer, 1976a).…”

Section: Methodsmentioning

confidence: 99%

Alkylation of ribose in RNA reacted with ethylnitrosourea at neutrality

Singer¹,

Kuśmierek²

1976

Biochemistry

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Ribose oxygens in TMV-RNA are ethylated by the carcinogen ethylnitrosourea in neutral aqueous solution (pH 6.1-7.3). 2'-O-ethyluridine, and 2'-O-ethylcytidine have been identified as reaction products. The four 2'-O-ethyl nucleosides are found in approximately equal amounts and the total extent of ribose alkylation is about 15% of the total ethylation. This finding, in conjunction with earlier results showing that all ring and phosphate oxygens can be ethylated, signifies that every oxygen in RNA or polyribonucleotides can react with ethylnitrosourea. The possible biological significance of ribose alkylation, resulting from chemical rather than enzymatic reaction, is discussed. The preparation of the new derivative 2'(3')-Oethylguanosine is described.

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“…Two dimensional paper chromatography of complete enzymatic hydrolysates of sycamore rRNA is reported in Fig.4 Spot 1 from both rRNAs was also made of a minor component which did not migrate in the system of Al-Arif and Sporn and behaved as N6-methyladenosine in solvent systems A, F, G and H [14,15]. Also, 1 M HCI hydrolysis of the material released a component migrating as N6-methyladenine in solvent D. Therefore, this compound was thought to be N6-methyladenosine for the 17-S rRNA.…”

Section: Two-dimensional Analysis Of Phosphatase Plus Phosphodiesreramentioning

confidence: 99%

Studies on the Methylation of Cytoplasmic Ribosomal RNA from Cultured Higher Plant Cells

Cecchini

Miassod

1979

European Journal of Biochemistry

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The methylation of cytoplasmic ribosomal RNA of cultured sycamore cells (Acer pseudoplatanus L.) was investigated. Labelled 17-S and 26-S rRNA were prepared from cells that had been incubated with either [32P]phosphate, methionine. Ion-exchange resin chromatography of 0.3 M KOH or 1 M HCl hydrolysates and two-dimensional chromatographic analyses of phosphodiesterase plus phosphatase digests of 17-S and 26-S rRNA were performed.17-S and 26-S rRNA contain 49 and 91 methyl groups per molecule, respectively. These values were verified in several ways. The high degree of methylation of sycamore rRNA, particularly for the 26-S rRNA, contrasts with the situation in all other investigated organisms.Several methylated bases were identified. 7-Methylguanine and 5-methylcytosine both occur in 17-S and 26-S rRNA. "-Methyladenine and N6,Nh-dimethyladenine are restricted to the 17-S rRNA while 3-methyluracil and 1 -methyladenine occur in the 26-S rRNA. One hypermodified uridine was also tentatively identified in the small rRNA. In 17-S rRNA, there is one copy of 7-methylguanine, N6-methyladenine and hypermodified uridine and two copies of N6,N6-dimethyladenine. 3-Methyluracil, 1-methyladenine and 5-methylcytosine occur twice, twice and three times, respectively, in 26-S rRNA. 7-Methylguanine and 5-methylcytosine are only in submolar amounts in the 26-S and 17-S rRNA, respectively.There are 40 & 2 and 83 f 3 2'-O-methylriboses per 17-S and 26-S rRNA molecule, respectively. In addition to the four 2'-O-methylnucleosides, one 2'-O-methylpseudouridine is present in the 17-S rRNA. Several lines of evidence argue for a non-random distribution of the methylriboses. In particular, one and seven Nm-Nm-Np structures occur in the 17-S and 26-S rRNA, respectively.The data are discussed comparatively with the methylation pattern of Escherichia coli, yeast and HeLa cell rRNA.The methylation of specific parts of polynucleotide chains is one of the multiple molecular events altering the primary transcriptional product of ribosomal genes and leading to the formation of the mature cytoplasmic rRNA [l]. Initial reports on the methylation of Escherichia coli rRNA and animal cytoplasmic rRNA suggested that eukaryotic cytoplasmic rRNA and prokaryotic rRNA strongly differ with regard to their type of methylation, the former being essentially ribose-methylated and the latter base-methylated. Later, specific methylated bases were identified in E. coli [2], yeast [3] and HeLa cells [4-61. These data supported the rather new idea that base-methylation might be an ancestral process widely conserved in living cells whereas ribose-methylation might represent a less-primitive process that appeared essentially in eukaryotic cells during evolution [l].With regard to cytoplasmic rRNA of higher plants, only 2'-O-methylation had been investigated [7-91. There had been no report on their basemethylation. Additional data on plant cells were

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An analytical method for the separation of sugar-methylated ribonucleosides from base-methylated and nonmethylated ribonucleosides

Cited by 27 publications

References 15 publications

5-methylcytosine content in the vertebrate deoxyribonucleic acids: Species specificity

5-methylcytosine content in the vertebrate deoxyribonucleic acids: Species specificity

Alkylation of ribose in RNA reacted with ethylnitrosourea at neutrality

Studies on the Methylation of Cytoplasmic Ribosomal RNA from Cultured Higher Plant Cells

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