Evidence for an “Early Early” Vaccinia Virus-Induced Protein Which Causes a Density Change of Infected L-M Cells

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The density decrease of vaccinia virus-infected L-M cells observed in a Ficoll density gradient by 2 h postinfection was found to be dependent on RNA synthesis and protein synthesis but independent of DNA synthesis. Using low multiplicities of infection, the required RNA and protein species appeared to be synthesized before parental viral DNA became sensitive to DNase, i.e., while the bulk of input virus was still at the core stage of uncoating. To date only thymidine kinase and a vaccinia virus-specific cell surface antigen (as well as the putative uncoating protein) have been shown to be “early early” proteins, i.e., synthesized while parental viral DNA is still enclosed within the core. Both heat-and UV-inactivated virus failed to cause the cell density decrease. The need for a functioning viral genome implies that the required early early RNA and protein species are virus specific and not cell specific. Thus the protein leading to the density decrease of L-M cells, induced very early after infection with vaccinia virus, represents one of the first bits of viral genetic information expressed after infection. Since antibody-neutralized virus is still capable of causing the phenomenon of cell density decrease, the basis of neutralization of vaccinia virus by specific antibody must be other than by inhibiting early early transcription and/or translation.

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