Virus Recovery in Chicken Cells Tested with Rous Sarcoma Cell DNA

Hill, M.; Hillova, Jana

doi:10.1038/newbio237035a0

Cited by 141 publications

(39 citation statements)

References 26 publications

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“…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)(5)(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).…”

mentioning

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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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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“…Briefly, after being washed twice with 0.15 m NaCI -0.1 m EDTA, pH 8.0. the cells were lysed with 0.5% sodium dodecyl sulfate in a buffer containing 0.1 M NaCI, 0.05 M Tris-HCl, pH 8.0, 0.01 M EDTA, and 500 gg/ml pronase. The lysate was digested at 37 for 4-5 h. Then sodium perchlorate was added to a final concentration of 1 m, and the DNA was deproteinized at room temperature with chloroform-isoamylalcohoi, precipitated with alcohol, and further processed as already indicated [9]. The extraction of rat thymus DNA (used as carrier DNA in the calcium method) was reported elsewhere [9],…”

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confidence: 99%

“…51 of chicken cells derived from a fibroblast infected and transformed, in all probability, by one particle of Schmidt-Ruppin RSV, subgroup D (SR-RSV-D). The cloning conditions were described elsewhere [8], The DNA was extracted as described [9] with slight modifications. Briefly, after being washed twice with 0.15 m NaCI -0.1 m EDTA, pH 8.0. the cells were lysed with 0.5% sodium dodecyl sulfate in a buffer containing 0.1 M NaCI, 0.05 M Tris-HCl, pH 8.0, 0.01 M EDTA, and 500 gg/ml pronase.…”

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confidence: 99%

Infectivity of Rous Sarcoma Cell DNA: Comparison of Two Techniques of Transfection Assay

Hillova¹,

Hill²,

Goubin³

et al. 1975

Intervirology

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A DNA extracted from a clone of chicken cells transformed by the Schmidt-Ruppin strain of Rous sarcoma virus, subgroup D (SR-RSV-D), was assayed for infectivity by means of DEAE-dextran and calcium techniques. The calcium technique like the previously described DEAE-dextran procedure gave rise to viruses in transfection assays with both native and denatured (SI nuclease susceptible) DNAs. The efficiency of these transfection techniques with native DNA was compared and found to be about the same provided that with the calcium technique carrier DNA was used to complement DNA concentrations lower than 2.5 µg/ml.

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“…Those findings overcame the objection of a one-way transmission of information from DNA to RNA and clinched in the minds of all but a few contrarians the DNA provirus hypothesis (81). If there was any doubt left, it would have been purged by the demonstration of infectious DNA from RSV-transformed, non-virus-producing rat cells (87). The latter resulted from the infection of newborn rats by the Prague strain of RSV propagated in Czechoslovakia (88).…”

Section: Dna Provirus Hypothesismentioning

confidence: 99%

The early history of tumor virology: Rous, RIF, and RAV

Rubin

2011

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

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One hundred years ago Peyton Rous recovered a virus, now known as the Rous sarcoma virus (RSV), from a chicken sarcoma, which reproduced all aspects of the tumor on injection into closely related chickens. There followed recovery of causal viruses of tumors of different morphology from 4 more of 60 chicken tumors. Subsequent studies in chickens of the biology of the first RSV isolated moved slowly for 45 y until an assay of ectodermal pocks of the chorioallantoic membrane of chicken embryos was introduced. The inadequacies of that assay were resolved with the production of transformed foci in cultures of chicken fibroblasts. There followed a productive period on the dynamics of RSV infection. An avian leukosis virus (ALV) was found in some chicken embryos and named resistance-inducing factor (RIF) because it interferes with RSV. Its epidemiology in chickens is described. Another ALV was found in stocks of RSV and called Rous-associated virus (RAV). Cells preinfected with RAV interfere with RSV infection, but RSV does not produce infectious virus unless RAV is added during or after RSV infection. Intracellular RAV provides the infectious coat for the otherwise defective RSV. The coat determines the antigenicity, host range, and maturation rate of RSV. RSV particles carry reverse transcriptase, an enzyme that converts their RNA into DNA and allows integration into the cell's DNA, where it functions as a cellular gene. This was the bridge that joined the biological era to the molecular era. Its relation to oncogenes and human cancer is discussed.

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Virus Recovery in Chicken Cells Tested with Rous Sarcoma Cell DNA

Cited by 141 publications

References 26 publications

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

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

Infectivity of Rous Sarcoma Cell DNA: Comparison of Two Techniques of Transfection Assay

The early history of tumor virology: Rous, RIF, and RAV

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