Integration of Herpes Simplex Virus Type 1 DNA into the DNA of Growth-arrested BHK-21 Cells

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Abstract
The ability of HSV-1 [herpes simplex virus type 1] DNA to become associated with host cell DNA in an alkaline-stable form was demonstrated following infection of a ts baby hamster kidney growth mutant (ts BTN-1), at the non-permissive temperature (39.5.degree. C). After 8 h pre-incubation at 39.5.degree. C, ts BTN-1 cells infected at this temperature using m.o.i. [multiplicity of infection] ranging from 0.5-200 p.f.u.[plaque forming units]/cell failed to replicate virus DNA even though transport of input virus genomes to the nucleus was the same at both permissive and non-permissive temperatures. Virions containing 3H-labeled DNA were used to infect ts BTN-1 cells at 39.5.degree. 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 DNA. 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.