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J Virol. 1969 September; 4(3): 256-263
Copyright © 1969 American Society for Microbiology. All Rights Reserved.

Fate of Adenovirus Type 12 Genomes in Nonpermissive Cells

Virus Laboratories, The Children's Hospital of Philadelphia, School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania, 19146
The Wistar Institute of Anatomy and Biology, Philadelphia, Pennsylvania 19104

ABSTRACT

The fate of ³H-thymidine-labeled adenovirus type 12 deoxyribonucleic acid (DNA) was studied in Nil-2 cells of Syrian hamster origin. It was found that a substantial fraction of ³H-adenovirus type 12 DNA became degraded within 24 hr after infection and was released into the culture fluid. After infection of 5-bromodeoxyuridine (BUdR)-prelabeled cells with ³H-adenovirus type 12, viral DNA became readily separable from cellular DNA by equilibrium centrifugation in CsCl. Part of the viral radioactivity was found to shift gradually to the position of cellular DNA as time progressed after infection. When exponentially growing cells were exposed simultaneously to BUdR, 5-fluorodeoxyuridine, and ³H-adenovirus type 12, up to 50% of the viral radioactivity shifted within 24 hr from the density of viral DNA to that of cellular DNA after equilibrium centrifugation in CsCl. Upon denaturation of the cellular DNA, the isotope was preferentially found to be associated with the "heavy" strand which was synthesized after infection. Upon hybridization of the "heavy" and the "light" strands with sonically treated, denatured ³H-adenovirus type 12 DNA, small and nearly equal amounts of counts hybridized with both strands. The number of counts annealed was in a range similar to that of those annealed with the same amount of DNA derived from adenovirus type 12-transformed hamster cells. These results demonstrate that (i) a substantial proportion of the adsorbed virus becomes degraded within 24 hr; (ii) part of the degradation products is reutilized for cellular DNA synthesis; (iii) only a small fraction, mainly fragments, of viral DNA becomes integrated into both the newly synthesized and the parental strands of cellular DNA.

¹ Present address: Virologisches Institut der Universität Würzburg, 8700 Würzburg, Germany.