Chromatin as a template for RNA synthesis in vitro.

Groner, Yoram; Monroy, Gladys; Jacquet, Michel; Hurwitz, Jerard

doi:10.1073/pnas.72.1.194

Cited by 30 publications

(10 citation statements)

References 19 publications

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“…Groner et al. (39) found that the greatest part of the RNA synthesized in chromatin isolated from chicken leukemic cells was in the form of RNA-DNA hybrids.…”

Section: Discussionmentioning

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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“…Groner et al. (39) found that the greatest part of the RNA synthesized in chromatin isolated from chicken leukemic cells was in the form of RNA-DNA hybrids.…”

Section: Discussionmentioning

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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“…Endoge nous ribonuc1eases and RNA polymerases are among these activities. While some workers have reported preparations of chromatin with no detectable RNase activity (27,28), most chromatin preparations contain RNase capable of reducing RNA to fragments of 4 to. 5S.…”

Section: Measurements Of Total Template Capacitymentioning

confidence: 99%

Regulation of Gene Expression in Eucaryotes

O’Malley¹,

Towle²,

Schwartz³

1977

Annu. Rev. Genet.

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“…A closed complex is sensitive to heparin until a temperature-dependent conformational change occurs to form the open complex (Yarbrough et al, 1976;Carpousis and Gralla, 1980;Hansen and McClure, 1980). In eukaryotic systems, heparin inhibits RNA polymerase II initiation but not elongation (Groner et al, 1975) and inhibits assembly of the RNA polymerase III transcription complex (Logan and Ackerman, 1988).…”

Section: Adriamycinmentioning

confidence: 99%

Drug Inhibitors of RNA Polymerase II Transcription

Logan¹,

Zhang²,

Davis³

et al. 1989

DNA

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Transcription by RNA polymerase II occurs after formation of a transcription complex. This complex is assembled in stages by the interaction of transcription factors with the template and/or with each other. We report on the ability of six drugs to inhibit the assembly of the RNA polymerase II transcription complex. Assembly of the complex on the adenovirus major late promoter requires several transcription factors. The normal assembly process requires that the DNA first interact with TFIIA, then with TFIID, and finally with at least four additional transcription factors (one of which is RNA polymerase II). We observed that streptolydigin (10 micrograms/ml) inhibits association of ILA and IID, and at higher concentrations (100 micrograms/ml) inhibits that IIA/IID complex from binding to DNA. Streptovaricin (100 micrograms/ml) appears to inhibit the IIA/IID interaction with DNA and prevents reinitiation (at 500 micrograms/ml). Adriamycin (1 microgram/ml) inhibits the interaction of TFIID with the IIA/DNA complex and inhibits an additional event immediately prior to, or during, elongation. Daunorubicin may be an elongation inhibitor. Heparin at 10 micrograms/ml inhibits further assembly after the IIA/IID/DNA complex has formed, and at 100 micrograms/ml also inhibits a late event in the assembly process and blocks reinitiation. Rifamycin AF/013 (100 micrograms/ml) inhibits the early events necessary to form the IIA/IID/DNA complex and (at 10 micrograms/ml) an assembly event following formation of the IIA/IID/DNA complex. Therefore, these compounds should be useful as probes for further examination of the assembly process.

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Chromatin as a template for RNA synthesis in vitro.

Cited by 30 publications

References 19 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

Regulation of Gene Expression in Eucaryotes

Drug Inhibitors of RNA Polymerase II Transcription

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