Distribution of repetitious sequences in chick nuclear DNA

Tapiero, Haïm; Monier, Marie‐Noëlle; Shaool, D.; Harel, J

doi:10.1093/nar/1.2.309

Cited by 22 publications

(11 citation statements)

References 16 publications

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“…In all cases, after attaining apparent saturation levels, 30-35% of the input 125I-labeled ssDNA hybridized to large excess amounts of homologous cytRNA was rendered S1 nuclease resistant, as compared with 4-4.5% for bulk nDNAt [the results, not shown, were comparable to all of those previously published (7)(8)(9)(10)(11)(12)(13)(14)]. The basepaired structure of ssDNA'RNA hybrids was verified by other methods.…”

supporting

confidence: 79%

“…1. RESULTS In all cases, ssDNA isolated from human lymphoid cells had the same characteristics as ssDNA isolated from other cell systems (7)(8)(9)(10)(11)(12)(13)(14). After two successive steps of hydroxylapatite chromatography purification, it amounted to 1.2-1.4% of the total nDNA and was 96-98% digested by S1 nuclease, whereas only 2-3% of bulk double-stranded nDNA was degraded under the same conditions.…”

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confidence: 80%

“…Over the past few years, such a fraction has been isolated in our laboratory in the form of a minor single-stranded component (ssDNA). In all normal or malignant cell systems studied, from human, murine, or avian origin, ssDNA amounted to 1.5-2% of the total nuclear DNA (nDNA); most of the ssDNA could be reassociated with unique-copy nDNA and 30-35% hybridized with homologous cytoplasmic RNAs (7)(8)(9)(10)(11)(12)(13)(14). It was also shown that 70-0% of ssDNA from the various cell systems (9)(10)(11)(12)(13)(14) consists ofnon-selfreassociating DNA chains, which can be reassociated with nonrepetitive nDNA and contains nearly all the ssDNA sequences hybridizable with homologous mRNAs (10).…”

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confidence: 99%

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Manifold expression of new cellular genes in human lymphoid neoplasia.

Hanania¹,

Shaool²,

Poncy³

et al. 1981

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

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Previous studies performed in our laboratory have shown that a minor single-stranded DNA component (ssDNA) isolated from the bulk of double-stranded nuclear DNA (nDNA) of human, murine, or avian cells consisted mainly of active transcription sites. In the present work, ssDNA isolated from human lymphoid cells, either malignant or not amounted to 1.2-1.4% ofthe total nDNA. Thiswas labeled with 5I and utilized as a probe in RNA-driven iterative hybridization and cross-hybridization assays. In all cases of neoplasia studied, the same subfraction of ssDNA (equivalent to 10-12% of the total ssDNA) from malignant cells could be hybridized with cytoplasmic RNA from malignant cells, whatever their origin: acute lymphoid leukemia cells collected from blood of leukemic patients, local lymphosarcomas, or cells from two established strains grown in suspension, one Epstein-Barr virus positive (Raji strain), the other Epstein-Barr virus negative (Ramos strain). The majority ofthese tumor-specific DNA sequences could be reassociated with unique-

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confidence: 79%

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confidence: 80%

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confidence: 99%

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Manifold expression of new cellular genes in human lymphoid neoplasia.

Hanania¹,

Shaool²,

Poncy³

et al. 1981

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

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“…The modification of the method for hydroxyapatite chromatography used in these studies has been described (15 of the native nuclear DNA from leukemic chicken cells was isolated as ss-DNA. Similar proportions of ss-DNA had been previously found in other cells including chicken embryonic cells (11,15,19,20). Whatever its origin, the ss-DNA could be entirely degraded by DNase or Si nuclease and was resistant to RNase or alkaline treatment.…”

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confidence: 99%

Single-stranded DNA from oncornavirus-infected cells enriched in virus-specific DNA sequences

Leibovitch¹,

Tapiero²,

Harel³

1977

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

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We previously found that a minor fraction of single-stranded DNA (ss-DNA) isolated from native nuclear DNA of normal chicken embryonic cells and cells of other species hybridized with bulk nuclear DNA or cellular RNA in great excess. At least one-third of ss-DNA belonging to the nonrepetitious part of the cell genome could be hybridized to homologous RNAs. In the present work, similar results were obtained with ss-DNA from cells of chickens infected by avian myeloblastosis virus (AMV). To investigate whether this enrichment of ss-DNA in transcribed DNA sequences involves provirus DNA, radioactive AMV RNA and cDNA copies of AMV RNA were used. Most of the 70S AMV RNA hybridized much faster to ss-DNA from productively infected leukemic cells than to bulk DNA. cDNA, either double-stranded or single-stranded, made in the presence of actinomycin D hybridized to total nuclear DNA with similar kinetics. In contrast, about half of the double-stranded cDNA molecules hybridized 40-50 times faster to ss-DNA than to total DNA, indicating that only one of the provirus DNA strands seems to be present in ss-DNA. This was confirmed by the fact that relatively insignificant amounts of the ss-cDNA molecules made in the presence of actinomycin D could be annealed to ss-DNA as compared with bulk DNA. These results indicate that actively transcribed DNA sequences can be selectively distributed in the ss-DNA fraction, probably because of single strand breaks in the vicinity of transcription sites.The provirus theory of Temin (1), which postulates that the replication of RNA tumor viruses proceeds via DNA copies of viral RNA integrated in the host genome, has been substantiated by the discovery of successful transfection mediated by reverse transcriptase (2, 3), through an intermediate of viral DNA isolated from oncornavirus-transformed cells (4-6) and molecular hybridization between the viral genome and cell nucleic acids (7,8). In productively infected cells, the integrated provirus DNA appears to be more actively transcribed than most of the cellular genes (9).We have isolated, from the nDNA of various species, a minor fraction of single-stranded DNA (ss-DNA) and demonstrated that about one-third of the ss-DNA from cultured normal embryonic chicken cells (10) The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be-hereby marked "advertsement" in accordance with 18 U. S. C. §1734 solely to indicate this fact.

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“…Molecular hybridizations. Purification of tumor-specific DNA Purification of cell nuclei, extraction of nDNA, isolation of ssDNA by two successive steps of hydroxylapatite chromatography with phosphate buffer at pH 7.85 (Tapiero et al, 1976), labeling with 1251, hybridization of labeled DNA with cellular RNA in large excess and determination of hybridization kinetics, monitored by the use of SI nuclease, were performed as previously described (Hanania et al, 1981). For iterative hybridization assays, the fraction of DNA hybridized to RNA (hybDNA) was separated from the nonhybridized DNA (non-hybDNA) by centrifuging to equilibrium in Cs2SO4 gradient in the absence of SI nuclease treatment (Hanania et al, 1981).…”

Section: Cells and Tissuesmentioning

confidence: 99%

Common multigenic activation in different human neoplasias.

Hanania¹,

Shaool²,

Harel³

et al. 1983

The EMBO Journal

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It was previously shown that a common multigenic component, designated as tumor‐specific DNA (tsDNA), was transcriptionally active in human lymphoid neoplasias and only slightly active, if at all, in normal lymphoid cells, including the Priess cell line immortalized by Epstein‐Barr virus. In the Burkitt lymphoma‐derived Raji cell line, tsDNA corresponded to 2500‐3000 distinct transcription units, arbitrarily defined as encoding mRNA chains of 1000 kds each. In the present study, radioactive Raji cell tsDNA was isolated by a recycling procedure which eliminates transcribed DNA sequences common to both the Raji cell and the Priess cell, and was used as a probe for homologous transcripts. The major part of this probe could be hybridized to RNAs from all the human neoplasias studied: cultured cell lines derived from leukemias, malignant lymphomas or sarcomas, leukemic cells or solid tumors (sarcomas and carcinomas) recovered from patients. In contrast, only a minor portion of Raji cell tsDNA could be hybridized to RNAs from non‐malignant cells, normal human lymphoid cells or fibroblasts grown in culture, fetal and chorioplacental tissues. It is concluded that a common multigenic set is activated in a wide variety of, and perhaps in all, human neoplasias.

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Distribution of repetitious sequences in chick nuclear DNA

Cited by 22 publications

References 16 publications

Manifold expression of new cellular genes in human lymphoid neoplasia.

Manifold expression of new cellular genes in human lymphoid neoplasia.

Single-stranded DNA from oncornavirus-infected cells enriched in virus-specific DNA sequences

Common multigenic activation in different human neoplasias.

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