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Compartmentalization of cellular and viral DNAs in adenovirus type 5 infection as revealed by ultrastructuralin situ hybridization

Chromosome Research volume 2, pages 123–135 (1994)Cite this article

Abstract

We describe the respective distributions of cellular DNA and viral genomes in adenovirus type 5 (Ad5)-infected HeLa cells by means of electron microscopein situ hybridization using biotinylated Alu DNA and Ad5 DNA probes in a post-embedding technique. When hybridization was performed on sections of formaldehyde-fixed material, Alu elements were restricted to the cellular chromatin, irrespective of the stage of the infectious cycle, without any interpenetration within the virus-induced regions. Viral DNA was confined entirely to the latter. Similar topological distributions of cellular and viral DNA were obtained whenin situ hybridizations were performed at the optical level on methanol—acetone fixed cells. Such separation between cellular and viral DNA persisted under experimental conditions which partially loosened the nucleoproteins. In addition, the viral DNA of the different viral structures spread without interpenetration. On the other hand, the procedure did not dissociate the clusters of viruses which remained, as usual, closely surrounded by innumerable viral genomes. It also preserved the shapes, content (viral ssDNA + viral 72 kD protein) and the topological localization of the pleomorphic viral ssDNA accumulation sites. These data, which suggest that all viral DNAs are spatially linked in the infected nuclei, suggest a prominent role for the internal nuclear matrix in virus replication, assembly and maturation.

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  1. Institut de Recherches Scientifiques sur le Cancer CNRS, Biologie et Ultrastructure du Noyau, 7 rue Guy Môquet, BP 8, 94801, Villejuif Cedex, France

    S. Besse & F. Puvion-Dutilleul

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  1. S. Besse
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  2. F. Puvion-Dutilleul
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Besse, S., Puvion-Dutilleul, F. Compartmentalization of cellular and viral DNAs in adenovirus type 5 infection as revealed by ultrastructuralin situ hybridization. Chromosome Res 2, 123–135 (1994). https://doi.org/10.1007/BF01553491

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  • DOI: https://doi.org/10.1007/BF01553491

Key words

  • adenovirus infection
  • Alu elements
  • biotinylated probes
  • DNA localization
  • electron microscopy
  • in situ hybridization