Nuclear and cytoplasmic ribonucleic acids of calf thymus

Osawa, Syozo; Takata, Kenzo; Hotta, Yasuo

doi:10.1016/0006-3002(58)90473-6

Cited by 97 publications

(16 citation statements)

References 18 publications

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“…Studies on a nucleus-free system: For studies of leucine-l-C'4 incorporation into proteins of isolated nuclear ribonucleoprotein particles in a nucleus-free system, the nuclei were isolated in 0.25 Al sucrose-0.003 M MgCl2, and freed of contaminating cytoplasmic fragments by two washings in an incubation medium of the follow- 15,17 Tris was used as the extracting buffer in all subsequent experiments. Addition of either Ca+ + or Mg++ did not enhance extraction.…”

mentioning

confidence: 99%

Metabolism and Morphology of Ribonucleoprotein Particles From the Cell Nucleus of Lymphocytes

Frenster

Allfrey²,

Mirsky³

1960

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

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Incubation of isolated nuclei of calf thymus lymphocytes with C'4-labelled precursor compounds results in rapid labelling of the proteins and ribonucleic acids (RNA) of the nuclei. ' Some of the proteins and RNA can then be extracted from the nuclei by neutral buffer solutions.' This paper describes the differentiM ultracentrifugation of such extracts which results in the isolation of classes of ribonucleoprotein particles, each of which can be characterized by its ease of extraction, its sedimentability, its composition of protein and RNA, and its rate of intranuclear protein and RNA metabolism. Unlike cytoplasmic ribonucleoprotein particles which are potassium-dependent,2 these nuclear particles require a sodium environment for their metabolic activity in the nucleus; furthermore, unlike cytoplasmic particles,3' 4the intranuclear metabolism of these nuclear particles is resistant to preincubation of the nuclei with ribonuclease (RNAase) but sensitive to pre-incubation of the nuclei with deoxyribonuclease (DNAase).It is also shown here that after extraction from the cell nucleus, the isolated nuclear ribonucleoprotein particles remain capable of active protein and RNA metabolism in a nucleus-free system.Previous studies of the cell nucleus by electron microscopy have demonstrated the occurrence of discrete particles of 100-300 A diameter in the nucleolus,5-7 on the lateral loops of lampbrush chromosomes,8 on the chromosomal rings of Balbiani,9 in the blebs'0 and pore annuli"' 12 of the nuclear membrane, and throughout the nuclear sap.13 Particles of similar size are shown here to be extracted from the nuclei of calf thymus lymphocytes, and to be composed of sub-units of 15-25 A diameter. The heterogeneity of the RNA of the cell nucleus has been shown previously by radioautography of labelled nuclei,'4 by isolation of labelled nuclear RNA species,'5' 16 by determination of the base composition of isolated nuclear RNA species,17 and by histochemical staining of nuclear ribonucleoprotein species throughout the mitotic cycle.'8 Composition and metabolic heterogeneity is; also

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mentioning

confidence: 99%

Metabolism and Morphology of Ribonucleoprotein Particles From the Cell Nucleus of Lymphocytes

Frenster

Allfrey²,

Mirsky³

1960

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

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“…Elution was carried out with a continuous elution system of dilute HCI instead of the formic acid system of Osawa et al [4]. A continuously increasing concentration of eluent was obtained in a mixing flask, between 50 ml distilled water and 30 ml 0.02 M HC1, and then between 50 ml solution and 50 ml 0.1 M HCI.…”

Section: Chromatographic Determination Of 'Nuckotides' Of Radioactivementioning

confidence: 99%

Synthesis in vitro of Novel Nucleic‐Acid‐Like Polymers of Ehrlich Ascites Tumor Cells

Kagawa

1983

European Journal of Biochemistry

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1. When cytoplasmic particles isolated from Ehrlich ascites tumor cells were incubated with [14C]ribose, bases and inorganic phosphate in the presence of MgC12 and Tris/HCl (pH 7.0), radioactivity was incorporated into trichloroacetic-acid-insoluble materials.2. Radioactive products, extracted and purified by DEAE-cellulose column chromatography, were hydrolyzed in 1 M HCl at 100°C for 1 h and the resultant radioactive neutral sugar components were purified and identified as ribose, glucose and rhamnose by gas chromatography and mass spectrometry.3. When [14C]glucose was incubated in the same reaction system, radioactivity was also incorporated into trichloroacetic-acid-insoluble materials. Purified radioactive products were hydrolyzed in 1 M HCl at 100 "C for 1 h and the resultant radioactive neutral sugar components were also shown to be ribose, glucose and a little rhamnose, as was the case with the products synthesized from [14C]ribose.4. When ['4C]deoxyribose was incubated in the same reaction system, little incorporation was observed. 5.The radioactive products were incubated with venom phosphodiesterase and the digests were analyzed by Dowex 1 x 4 column chromatography. The results suggest that the radioactive products contained ribonucleotides and 'nucleotides' containing glucose (glucose 'nucleotides').6. When the radioactive products were incubated with both venom phosphodiesterase and alkaline phosphatase, ribonucleosides and glucose 'nucleosides' were detected. In Dowex 1x4 (borate form) chromatography, ribonucleosides were eluted coincidentally with ribonucleoside markers (Cyd, Ado, Urd and Guo), whereas the elution profiles of glucose 'nucleosides' were totally different from those of ribonucleosides.

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“…RNA reparation extracted with Tris (pH 7.4)-phenol was hydrolyzed to mononucleotides by incubation with 0.5 N KOH at 37°C for 18h. According to the method of Osawa et al [21], the ribonucleotides were fractionated on formate type Dowex-1 ion exchange resin column chromatography by elution with gradient of formic acid. Then T radioactivity and A260 or A280 of each fraction were measured.…”

Section: Preparation Of Nucleic Acids and Fractionation Of Mononucleomentioning

confidence: 99%

Accumulation and retention of Tritium (tritiated water) inRhodopseudomonas spheroides under aerobic condition

Inomata

Higuchi

1982

Radiat Environ Biophys

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Summary. Nonsulfur purple photosynthetic bacteria, Rhodopseudomonas spheroides cells grown aerobically in the dark were incubated with medium containing tritiated water (THO) under growth and no-growth conditions. R value defined as the ratio of specific activity (T/H) of tritium (T) incorporated into cell materials to T in medium was about 0.5 in growing cells and 0.2 in nongrowing cells regardless of THO-concentration in medium. From the fractionation of these lyophilized cells by modified Schneider method, the distribution of volatile T (i.e., exchangeable T) in cold perchloric acid (PCA)-soluble fraction, nonexchangeable T of small molecules in cold PCA-and ether-ethanol-soluble fractions and nonexchangeable T of macromolecules in hot PCA-and alkali-soluble fractions was 16 : 17 : 67 in growing cells and 38 : 24 : 38 in nongrowing cells. Moreover, the nucleic acids were extracted by phenol method from the cells incubated with THO and fractionated on methylated albumin-coated column chromatography. R value of nonexchangeable H in DNA, sRNA, and rRNA was 0.4-0.6 in growing cells and 0.06-0.13 in nongrowing cells. R value of nonexchangeable H in ribo-and deoxyribo-mononucleotides hydrolyzed by treatment with either KOH or DNase and phosphodiesterase was 0.38-0.84 in growing cells. Average R value of nonexchangeable H in nucleic acids and these mononucleotides in growing cells were similar to that of total H in the lyophilized cells. From the study of retention of T in the cells, radioactivity of T of long-lived component decreased at the biological half life of 69 h in nongrowing cells and at rate of 0.4 n (n: number of cell division) in growing cells. The retention of bound T in nucleic acids of nongrowing cells was considerable, i.e., 100, 78, and 86% remaining after 16 h in DNA, sRNA, and rRNA, respectively, for 76% in the overall cells. Decrease of activity of nonexchangeable T bound to DNA in nongrowing cells was expected only the cell death.

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Nuclear and cytoplasmic ribonucleic acids of calf thymus

Cited by 97 publications

References 18 publications

Metabolism and Morphology of Ribonucleoprotein Particles From the Cell Nucleus of Lymphocytes

Metabolism and Morphology of Ribonucleoprotein Particles From the Cell Nucleus of Lymphocytes

Synthesis in vitro of Novel Nucleic‐Acid‐Like Polymers of Ehrlich Ascites Tumor Cells

Accumulation and retention of Tritium (tritiated water) inRhodopseudomonas spheroides under aerobic condition

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