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Open AccessJournal ArticleDOI

State of Adenovirus 2 Deoxyribonucleic Acid in the Nucleus and Its Mode of Transcription: Studies with Isolated Viral Deoxyribonucleic Acid-Protein Complexes and Isolated Nuclei

Robert D. Wallace, +1 more
- 01 Apr 1972 - 
- Vol. 9, Iss: 4, pp 627-635
TLDR
Newly replicated adenovirus 2 deoxyribonucleic acid (DNA) can be isolated from the nucleus of HeLa cells by a gentle lysis procedure as a fairly homogeneous complex with a sedimentation of 73S and the synthesis of virus-specific RNA by the isolated nuclei is strongly inhibited by low doses of alpha-amanitine.
Abstract
Newly replicated adenovirus 2 deoxyribonucleic acid (DNA) can be isolated from the nucleus of HeLa cells by a gentle lysis procedure as a fairly homogeneous complex with a sedimentation of 73S. The viral DNA complex can be prepared completely free from host cell DNA. The viral complex is slightly active in ribonucleic acid (RNA) synthesis in vitro. Treatment of the complex with Pronase and sodium dodecyl sulfate converts the DNA to a form which sediments at 43S. Nuclei isolated from adeno-infected cells synthesize high-molecular-weight virus-specific RNA in vitro. Optimal RNA synthesis requires a divalent cation, preferentially manganese, and relatively high salt concentrations. The synthesis of virus-specific RNA by the isolated nuclei is strongly inhibited by low doses of alpha-amanitine. The latter experimental result is discussed in terms of the polymerase used to transcribe the adenovirus DNA in vivo.

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Citations
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Journal ArticleDOI

An amazing sequence arrangement at the 5′ ends of adenovirus 2 messenger RNA

TL;DR: Findings imply a new mechanism for the biosynthesis of Ad2 mRNA in mammalian cells which is complementary to sequences within the Ad2 genome which are remote from the DNA from which the main coding sequence of each mRNA is transcribed.
Journal ArticleDOI

Selective and accurate initiation of transcription at the ad2 major late promotor in a soluble system dependent on purified rna polymerase ii and dna

TL;DR: Specific transcription initiation at the late promoter is observed with extracts derived from either virus-infected or uninfected KB cells and with class II RNA polymerases isolated from either human calf, murine or amphibian cells.
Journal ArticleDOI

Promoters and heterogeneous 5' termini of the messenger RNAs of adenovirus serotype 2.

TL;DR: A model for Ad2 cap site recognition is proposed in which RNA polymerase scans short DNA segments for purine starts, and is discussed in the context of EIV mRNAs of Ad2 and Ad5, which differ in their promoter region DNA sequence.
Journal ArticleDOI

Transcripts from the adenovirus-2 major late promoter yield a single early family of 3′ coterminal mRNAs and five late families

TL;DR: It is shown that a major event in the Ad-2 early-late switch is a novel form of control which activates L2-L5 mRNA production, and implies that precisely the same transcription start site is utilized for early L1 mRNA synthesis as is used during the late stage for L1-L 5 late mRNA synthesis.
Journal ArticleDOI

A new species of virus-coded low molecular weight RNA from cells infected with adenovirus type 2

TL;DR: A virus-coded low molecular weight RNA (5.2S), which migrates slightly faster on polyacrylamide gels than the well characterized adenovirus-specific 5.5S RNA, has been isolated from cells infected with adenOVirus type 2.
References
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Journal ArticleDOI

Multiple Forms of DNA-dependent RNA Polymerase in Eukaryotic Organisms

TL;DR: Three distinct RNA polymerase activities have been isolated from developing sea urchin embryos, and in rat liver nuclei there are two RNA polymerases activities.
Journal ArticleDOI

Specific Inhibition of Nuclear RNA Polymerase II by α-Amanitin

TL;DR: α-Amanitin, a toxic substance from the mushroom Amanita phalloides, is a potent inhibitor of DNA-dependent RNA polymerase II (the nucleoplasmic form) from sea urchin, rat liver, and calf thymus.
Journal ArticleDOI

Specific Nucleolar and Nucleoplasmic RNA Polymerases

TL;DR: It is concluded that the RNA polymerases are specifically localized within the nucleus and may, therefore, play specific roles in the regulation of genetic transcription.
Journal ArticleDOI

Biochemical studies on adenovirus multiplication: IV. Isolation, purification, and chemical analysis of adenovirus☆

TL;DR: Type 4 adenovirus prepared by the above procedure had a similar chemical composition and DNA base ratios as type 2 adenova, and evidence for the purity of the virus and for the double-stranded nature of viral DNA is discussed.
Journal ArticleDOI

The Folded Genome of Escherichia coli Isolated in a Protein-DNA-RNA Complex

TL;DR: The DNA unfolds after exposure to RNase or heat; this indicates that an RNA component of the complex is involved in stabilizing the structure.
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