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Nuclear Localization and Covalent Linkage of Infective Virus DNA to Chromosomal DNA of Non-producer Rous Sarcoma Cells

Dominique Coulaud

^* Department of Cellular and Molecular Biology and Equipe de Recherche N^o 148 du C.N.R.S., Institute of Cancerology and Immunogenetics, Villejuif, France
Laboratory for Electron Microscopy, Institute Gustave-Roussy, Villejuif, France

The nuclei of Rous sarcoma cells were prepared from an established line of non-producer rat XC cells transformed with a Prague strain of Rous sarcoma virus (PR-RSV). Electron microscopic examination of the nuclear pellet showed a slight contamination with cytoplasmic debris and an absence of mitochondria. The DNA samples extracted from isolated nuclei and from whole XC cells were both assayed for infectivity in chicken cell cultures and found to contain about the same number of infective units per unit weight of DNA. Furthermore, the DNA from whole XC cells was set free by alkali and sedimented through an alkaline glycerol gradient in order to separate cellular DNA species according to sedimentation velocity. Under these conditions the infective RSV DNA consistently sedimented with the chromosomal 110S DNA and thus behaved as if covalently linked to the chromosomal DNA of XC cells. These results show that the infective virus DNA of non-producer RSV-transformed cells is carried in these cells as an integral part of the cellular chromosome.

Received 23 November 1973; accepted 17 January 1974.