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J Gen Virol 44 (1979), 657-667; DOI 10.1099/0022-1317-44-3-657
© 1979 Society for General Microbiology
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Integration of Herpes Simplex Virus Type 1 DNA into the DNA of Growth-arrested BHK-21 Cells

Kazuo Yanagi, Mark G. Rush and Ken Biegeleisen

Department of Biochemistry, New York University School of Medicine, 550 First Avenue, New York, New York 10016, U.S.A.

The ability of HSV-1 DNA to become associated with host cell DNA in an alkaline-stable form has been demonstrated following infection of a ts baby hamster kidney growth mutant (ts BTN-1), at the non-permissive temperature (39.5 °C). After 8 h pre-incubation at 39.5 °C, ts BTN-1 cells infected at this temperature using m.o.i. ranging from 0.5 to 200 p.f.u./cell fail to replicate virus DNA even though transport of input virus genomes to the nucleus is the same at both permissive and non-permissive temperatures.

Virions containing 3H-labelled DNA were used to infect ts BTN-1 cells at 39.5 °C, and the total cellular DNA isolated from these cells was resolved into host and virus material by repeated CsCl equilibrium gradient centrifugation. A significant amount of the input radioactivity was found as a distinct band in the host region in both neutral and alkaline CsCl gradients, strongly suggesting a covalent association between host and virus DNAs. Evidence for this association was strengthened by demonstrating that radioactive material (virus DNA) banding in the host region of CsCl gradients could be driven towards the density expected for virus DNA following degradation of the putative hybrid molecules by shearing.

Received 24 October 1978; accepted 22 February 1979.




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Copyright © 1979 by the Society for General Microbiology.