High molecular weight nonribosomal-type nuclear RNA and cytoplasmic messenger RNA in HeLa cells.

Houssais, Jean‐François; Attardi, Giuseppe

doi:10.1073/pnas.56.2.616

Cited by 86 publications

(21 citation statements)

References 14 publications

(3 reference statements)

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“…Since the discovery of the similar DNA-like base compositions of HnRNA and mRNA, the hypothesis has been considered (22)(23)(24)(25) that mRNA is fashioned from a higher molecular weight nuclear precursor. Decisive experiments for or against this proposal are still lacking.…”

Section: Methodsmentioning

confidence: 99%

An Adenylic Acid-Rich Sequence in Messenger RNA of HeLa Cells and Its Possible Relationship to Reiterated Sites in DNA

Darnell

Wall

Tushinski

1971

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

367

111

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Messenger RNA from HeLa cells contains, as part of the polynucleotide chain, RNase-resistant sequences that are labeled by adenosine but not by uridine. Heterogeneous nuclear RNA also contains adenylate-rich RNase-resistant regions, but in lower proportion than messenger RNA. Hybridization to DNA of 32P-labeled messenger RNA reveals that some of the adenylate-rich region is included in the rapidly-hybridizing fraction.

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Section: Methodsmentioning

confidence: 99%

An Adenylic Acid-Rich Sequence in Messenger RNA of HeLa Cells and Its Possible Relationship to Reiterated Sites in DNA

Darnell

Wall

Tushinski

1971

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

367

111

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“…In addition to the ribosomal RNA species and their large precursors, one can see in the sedimentation profile of nuclear RNA small amounts of 48 RNA, and a polydisperse RNA with sedimentation constants between 6S and more than 50S. The latter material presumably represents, at least in part, the heterogeneous nonribosomal type nuclear RNA described in HeLa cells (34) and other animal cells (13,35,36).…”

Section: Sedimentation Profile Of Rna From Isolated Chromosomesmentioning

confidence: 99%

Isolation of Metaphase Chromosomes From Hela Cells

Huberman

Attardi

1966

The Journal of Cell Biology

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The authors have developed a method for large-scale isolation of metaphase chromosomes from HeLa cells. The distinguishing feature of this method is the use of a pH sufficiently low (about 3) to stabilize the chromosomes against mechanical damage. Many milligrams of fairly pure, morphologically intact chromosomes can be isolated in 8 hr or less of total working time. The isolated chromosomes contain about 2.0 mg of acid-soluble protein, 2.7 mg of acid-insoluble protein and 0.66 mg of R N A for each milligram of DNA. The R N A bound to the isolated chromosomes consists mainly of ribosomal RNA, but there is also a significant amount of 45S RNA. I N T R O D U C T I O NMany possible biochemical and biophysical approaches to the study of chromosomes in higher organisms have been hindered, until recently, by the lack of suitable procedures for large-scale isolation of chromosomes. Although the methods for isolation of interphase chromosomes, or "chromatin," which have been developed in recent years (1, 2) are satisfactory for certain purposes, a definite need still exists for a procedure which will allow large-scale isolation of morphologically intact metaphase chromosomes. Metaphase chromosomes are an indispensable complement to interphase chromosomes for the general study of chromosome structure. In addition, metaphase chromosomes have the unique advantage of being so condensed that they can be distinguished microscopically both from each other and from contaminating nonchromosomal material. Consequently, one is not limited to studying the average properties of all chromosomes; one can also examine single types of chromosomes.According to our experience, in the isolation of metaphase chromosomes by most previously published methods (3-5), morphological damage to some of the chromosomes cannot be avoided and only partial purification of the chromosomes from cell debris can be achieved. We report here a method for the rapid preparation, in milligram quantities, of fairly pure, morphologically intact metaphase chromosomes from HeLa cells. We also report the results of studies on the chemical composition of isolated chromosomes. M A T E R I A L A N D M E T H O D S Cultivation of CellsHeLa $3 cells (6) were grown in suspension culture in a modified Eagle's medium (7) supplemented with 5% calf serum. For accumulation of metaphase cells, partial synchrony was induced by lowering the culture temperature to 4°C for 1 hr and then returning it to 37°C (8). Ten to 11 hr later, colchicine was added to a final concentration of 0.5 to 1 X 10 -5 M. The cells were harvested by centrifugation 9 to 10 hr after colchicine addition and washed 3 times in 0.137M NaC1, 0.005M KC1, 0.007M NaH2PO4, 0.025 m Tris, pH 7.4. This procedure routinely produced about 30% metaphase cells. 95 on

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“…The second part is the informative part which is destined to become messenger RNA. The two parts together, that is the initial whole molecule, he proposes, is the heavy heterogeneous nuclear RNA reported by several laboratories (41)(42)(43). Various regulatory proteins interact with sites on the DNA from which the non-informative part of the RNA is transcribed.…”

mentioning

confidence: 99%

Molecular events basic to cellular radiation response. Progress report

Kolodny¹

1974

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High molecular weight nonribosomal-type nuclear RNA and cytoplasmic messenger RNA in HeLa cells.

Cited by 86 publications

References 14 publications

An Adenylic Acid-Rich Sequence in Messenger RNA of HeLa Cells and Its Possible Relationship to Reiterated Sites in DNA

An Adenylic Acid-Rich Sequence in Messenger RNA of HeLa Cells and Its Possible Relationship to Reiterated Sites in DNA

Isolation of Metaphase Chromosomes From Hela Cells

Molecular events basic to cellular radiation response. Progress report

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