Method for determination of the methylated constituents of transfer ribonucleic acid

Munns, Theodore W.; Podratz, Karl C.; Katzman, Philip A.

doi:10.1021/bi00718a026

Cited by 41 publications

(23 citation statements)

References 31 publications

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“…9; unpublished). DNA was isolated as described above (yielding 35,000-100,000 cpm per ,g of DNA) and hydrolyzed with 88% (wt/vol) formic acid at 150°C for 30 min (9,14). The released purine and pyrimidine bases along with standards were separated by descending paper chromatography in 2-propanol/concentrated HCl/H2O (680:176:144, vol/vol).…”

mentioning

confidence: 99%

Methylation of Herpesvirus saimiri DNA in lymphoid tumor cell lines

Desrosiers

Mulder

Fleckenstein

1979

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

171

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Several continuous lymphoid cell lines have been established from tumors induced by Herpesvirus saimiri . At least a portion of the viral DNA in the marmoset lymphoid cell line 1670, which does not produce detectable virus, is present as covalently closed circular episomal DNA. The use of restriction endonuclease digestion, transfer to nitrocellulose filters, and hybridization of the virus-specific DNA has produced strong evidence that viral DNA sequences present in total 1670 cell DNA and in isolated episomes are extensively methylated. The restriction endonuclease Hpa II has the same recognition sequence as Msp I but, unlike Msp I, fails to cleave when the C of the C-G dinucleotide is methylated. Viral DNA sequences of 1670 cells are refractory to cleavage by Hpa II but not Msp I; greater than 80% of the Hpa II cleavage sites appear to be methylated. Similarly, viral DNA sequences of 1670 cells are refractory to cleavage by Sma I (C-C-C-G-G-G) and Sac II (C-C-G-C-G-G) but not Sac I, Pvu II, or Pst I, which lack the dinucleotide C-G in their recognition sequences. Methylation of mammalian DNA has been previously found exclusively at C residues in the dinucleotide C-G. H. saimiri DNA sequences of another nonproducer cell line, 70N2, also appeared to be extensively methylated, but analysis of total cell DNA extracted from three virus-producing lymphoid lines revealed no evidence of methylation of viral DNA sequences. It remains to be seen if methylation of viral DNA plays a role in the lack of complete expression of H. saimiri genome information in nonproducing lymphoid cell lines.

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mentioning

confidence: 99%

Methylation of Herpesvirus saimiri DNA in lymphoid tumor cell lines

Desrosiers

Mulder

Fleckenstein

1979

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

171

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“…These values are equivalent to the amounts of riboTp, m Gp and m Ap in tRNA from other strains of E. coli (11,17,18). The identity of riboTp was confirmed by the mass number of the nucleoside, 258 daltons, as determined by mass spectroscopy of the nucleotide that had been eluted from Cellulose scrapings of thin layer chromatography plates.…”

Section: Othersmentioning

confidence: 89%

Utilization of an Escherichia coli mutant for carbon-13 enrichment of tRNA for NMR studies

Agris¹,

Fujiwara²,

Schmidt³

et al. 1975

Nucl Acids Res

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The enrichment of tRNA at specific sites with carbon-13 has been accomplished in vivo using a mutant of Escherichia coli. A relaxed strain of E. coli auxotrophic for methionine was grown in a specifically defined medium supplemented with either [14C] or [13C]-methyl labeled methionine. Cells were collected at the end of the log-phase of growth and tRNA was extracted. Analysis of the radioactivity of the [14C]-labeled tRNA established an incorporation ratio of three labeled carbons per tRNA molecule. Incorporation of the [14C]-label in vivo was confined to the methylation of nucleotides as determined by thin layer chromatography of nucleotides resulting from a ribonuclease digestion of [14C]-labeled tRNA. The carbon-13 NMR spectrum of [13C]-enriched tRNA indicated a similar degree of incorporation into the methylated nucleotides by the substantial enhancement of [13C]-methyl NMR signals only. Assignment of signals has been made for the methyl groups of ribothymidine and N7-methylguanosine in E. coli tRNA.

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“…The volumes of the samples were brought back to 20 ul after evaporation to dryness and chromatographed by thin-layer chromatography for 4 h on silica gel plates with ethyl acetate, methanol, water, 88% formic acid (100:25:20:1; vol/vol). Known methylated and nonmethylated bases were used as controls (14).…”

Section: Methodsmentioning

confidence: 99%

Inhibition of viral RNA methylation in herpes simplex virus type 1-infected cells by 5' S-isobutyl-adenosine

Jacquemont¹,

Huppert²

1977

J Virol

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5' S-isobutyl-adenosine (SIBA), a structural analogue of S-adenosylhomocysteine, reversibly blocks the multiplication of herpes simplex type 1 virus. In the presence of SIBA, viral protein synthesis is inhibited. After removing SIBA the synthesis of proteins starts rapidly again. The new polypeptides are mainly alpha proteins (Honess and Roizman, J. Virol. 14:8-19, 1974,), normally the first to be synthesized after infection. The rapid synthesis of proteins after release of inhibition seems to be directed by mRNA formed in the presence of SIBA as indicated by experiments using actinomycin D but which was undermethylated as shown by analysis of methyl groups on RNA. SIBA inhibits the methylation of mRNA and especially that of the 5' cap. Capping of mRNA thus seems to be essential for efficient translation. The analogue affected various methylations to different extents.

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Method for determination of the methylated constituents of transfer ribonucleic acid

Cited by 41 publications

References 31 publications

Methylation of Herpesvirus saimiri DNA in lymphoid tumor cell lines

Methylation of Herpesvirus saimiri DNA in lymphoid tumor cell lines

Utilization of an Escherichia coli mutant for carbon-13 enrichment of tRNA for NMR studies

Inhibition of viral RNA methylation in herpes simplex virus type 1-infected cells by 5' S-isobutyl-adenosine

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