Intracellular forms of adenovirus deoxyribonucleic acid. I. Evidence for a deoxyribonucleic acid-protein complex in baby hamster kidney cells infected with adenovirus type 12.

Journal of virology ·Vol. 9 ·No. 2 ·1972-02-00 ·Pages 297-308

Doerfler W, Lundholm U, Hirsch-Kauffmann M

Abstract

The total intracellular deoxyribonucleic acid (DNA) from baby hamster kidney cells abortively infected with (3)H-adenovirus type 12 was analyzed in dye-buoyant density gradients. Between 10 and 20% of the cell-associated radioactivity derived from viral DNA bands in a density position which is 0.043 to 0.085 g/cm(3) higher than that of viral DNA extracted from purified virions. The DNA in the high-density region (HP-fraction) is almost completely absent when DNA, ribonucleic acid (RNA) or protein synthesis is chemically inhibited in separate experiments. The HP-fraction is not found when the virus does not adsorb to and enter the cell. The DNA in the HP-fraction appears as early as 2 hr after inoculation. At 2 hr after infection, the HP-fraction is present both in the nucleus and the cytoplasm. This DNA hybridizes exclusively with viral DNA and sediments at approximately the same rate in both neutral and alkaline sucrose density gradients. Electron microscopy has revealed no circular DNA molecules in this fraction. Evidence indicates that the viral DNA in the HP-fraction exists in a complex with protein and possibly RNA. The protein component of the complex is resistant to enzymatic digestion, whereas the complex is susceptible to ribonuclease treatment. Digestion with deoxyribonuclease reduces the amount of DNA found in the HP-fraction. The structure and biological function of this complex are currently being investigated.

MeSH Terms

Adenoviridae/analysis Adsorption Animals Carbon Isotopes Carcinoma Cell Line Cell Nucleus/analysis Centrifugation, Density Gradient Centrifugation, Zonal Cricetinae Cytoplasm/analysis DNA, Viral/analysis Deoxyribonucleases Electrophoresis, Disc Glucosamine/analysis Humans Kidney Mouth Neoplasms Nucleic Acid Hybridization Pronase Protein Binding Ribonucleases Tritium Viral Proteins/analysis Virus Cultivation

Chemicals

Carbon Isotopes DNA, Viral Viral Proteins Tritium Deoxyribonucleases Ribonucleases Pronase Glucosamine

Authors & Affiliations

3 authors, click to expand affiliations / ORCID

Doerfler W

Lundholm U

Hirsch-Kauffmann M

References (33)

33 references, click to expand

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Article Info

Journal

Journal of virology

Abbr.

J Virol

ISSN

0022-538X

DOI

10.1128/JVI.9.2.297-308.1972

Published

1972-02-00

Pages

297-308

Language

English

Region

United States

NLM ID

0113724

PMCID

PMC356297

Subset

External Links

PubMed source DOI: 10.1128/JVI.9.2.297-308.1972 PMC Full Text View journal details

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Intracellular forms of adenovirus deoxyribonucleic acid. I. Evidence for a deoxyribonucleic acid-protein complex in baby hamster kidney cells infected with adenovirus type 12.

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