Chromosomal ribonucleic acid of rat ascites cells

Dahmus, Michael E.; McConnell, Dorothy

doi:10.1021/bi00832a031

Cited by 77 publications

(25 citation statements)

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“…The experimental procedures were based on the methods of Sabatani et al (1962), Muramatsu, Smetana and , DiGirolamo, Henshaw and Hiatt (1964), Dahmus and McConnell (1969) and Culp and Brown (1970). The rats were killed by cervical dislocation and the excised tumour tissue, freed from any necrotic material, was collected in medium A (0-25 mol/l sucrose, 3 mmol/l CaCl2, 50 mmol/l Tris-HCl buffer, pH 7-6; DiGirolamo et al., 1964) at 00.…”

Section: Methodsmentioning

confidence: 99%

“…A portion of the suspension was treated with ultrasonic vibrations as described above, and then centrifuged for 5 min at 2500g. Chromatin was recovered from the supernatant fraction by centrifugation for 10 min at 10,000g, and purified as described by Dahmus and McConnell (1969). The nucleolar pellet was resuspended in RSB to a volume of 5-5 ml and 5 ml of this suspension mixed with an equal volume of medium C (1-76 mol/l sucrose, 6-6 mmol/l CaCl2 and 20 mmol/l Tris-HCl buffer, pH 7-6).…”

Section: Methodsmentioning

confidence: 99%

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Intranuclear Distribution of the Inducing Metal in Primary Rhabdomyosarcomata induced in the Rat by Nickel, Cobalt and Cadmium

Webb¹,

Heath²,

Hopkins³

1972

Br J Cancer

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Intranuclear Distribution of the Inducing Metal in Primary Rhabdomyosarcomata induced in the Rat by Nickel, Cobalt and Cadmium

Webb¹,

Heath²,

Hopkins³

1972

Br J Cancer

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“…RNA polymerase assay: RNA polymerase was assayed as previously described [2]. Ascites DNA was prepared following the method of Dahmus and McConnell [4] .…”

Section: North-holland Publishing Company -Amsterdammentioning

confidence: 99%

Multiple froms of a eukaryotic RNA polymerase II stimulation factor

Dahmus¹

1976

FEBS Letters

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“…Chromosomal RNA (eRNA) is a small nuclear RNA that comprises a significant fraction of the total nuclear RNA in all higher eukaryotes investigated (1)(2)(3). It hybridizes to the repetitive DNA sequences (4,5) and is distinguished by a high content of dihydropyrimidine (6).…”

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confidence: 99%

A Partial Sequence of Nuclear Events in Regenerating Rat Liver

Mayfield

Bonner

1972

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

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When two lobes of the liver of the adult rat are removed, the cells of the remaining lobe are aroused to renewed cell division. We have studied the early events in such regeneration. The first observable response to such partial hepatectomy is the production in the liver nuclei of rapidly-labeled high-molecular-weight RNA of sequences not produced by normal liver. This is followed, with a lag of about 1 hr, by the appearance of increased (above normal) amounts of chromosomal RNA, again of sequences not produced by normal liver. With a lag of another hour, the template activity in support of RNA synthesis of the liver chromatin increases substantially. These events occur before initiation of DNA synthesis in the cells of thAe regenerating liver.Chromosomal RNA (eRNA) is a small nuclear RNA that comprises a significant fraction of the total nuclear RNA in all higher eukaryotes investigated (1-3). It hybridizes to the repetitive DNA sequences (4, 5) and is distinguished by a high content of dihydropyrimidine (6). Several experiments have suggested that cRNA is involved in the processes of gene activation (7,3). In addition, it has recently been shown that cRNA and high-molecular-weight rapidly-labeled nuclear RNA (Hn RNA) share a large proportion of their nucleotide sequences, a result that strongly suggests a precursor-product relationship between them (5). To shed further light on these matters, we have investigated the temporal relationships between Hn RNA, cRNA, and template activity in regenerating rat liver. MATERIALS AND METHODSRegenerating Livers. The livers of 200-400 g male, albino Sprague-Dawley rats were caused to regenerate by surgically removing their two largest lobes (9). All operations were performed on rats anesthetized with ether, and timed so that the regenerating livers were harvested between 6 p.m. and 10 p.m. to minimize circadian effects. The rats were decapitated and their livers were quickly frozen in liquid nitrogen.Chromatin. 20-40 g of frozen livers in saline-EDTA buffer (75 mM NaCl-24 mM EDTA, pH 8) were homogenized in a Waring blendor (85 V for 10 sec, 30 V for 3 min). This solution was then filtered through Miracloth (Chicopee Manufacturing Co., Milltown, N.J.) and centrifuged at 3500 rpm in the Sorvall centrifuge (SS-34 rotor) for 10 min. Crude chromatin was made from the pellet by three or four cycles of glass-teflon homogenization and centrifugation at 10,000 rpm (SS-34 rotor) for 10 min in 0.01 M Tris buffer (pH 8) (8). Chromatin for the template assays was purified by centrifugation through 1.7 M sucrose. The pellet was washed once by resuspension in 0.01 M Tris buffer (pH 8) and centrifugation at 10,000 rpm for 10 min. It was then sheared in a Virtis homogenizer at 45 V for 90 sec at a concentration of 10 A260/ ml. This solution was centrifuged for 30 min at 10,000 rpm in the Sorvall centrifuge and the supernatant was carefully removed and frozen in liquid nitrogen (8).Preparation of Chromosomal RNA. Chromosomal RNA was prepared by one of two previously described method...

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Chromosomal ribonucleic acid of rat ascites cells

Cited by 77 publications

References 22 publications

Intranuclear Distribution of the Inducing Metal in Primary Rhabdomyosarcomata induced in the Rat by Nickel, Cobalt and Cadmium

Intranuclear Distribution of the Inducing Metal in Primary Rhabdomyosarcomata induced in the Rat by Nickel, Cobalt and Cadmium

Multiple froms of a eukaryotic RNA polymerase II stimulation factor

A Partial Sequence of Nuclear Events in Regenerating Rat Liver

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