Insertional inactivation of the large subunit of ribonucleotide reductase encoded by vaccinia virus is associated with reduced virulence in vivo.

Abstract

To assess whether a fully functional VV ribonucleotide reductase enzyme is required during both in vitro and in vivo replication of VV, three mutant viruses were constructed by marker transfer techniques: M1 lambda, an M1 insertion mutant; TK-, an insertion mutant of the VV thymidine kinase (tk) gene; and M1 lambda/TK-, a double mutant. Extracts of cells infected with the M1 lambda or M1 lambda/TK- mutant viruses were assayed for ribonucleotide reductase activity and it was found that insertional inactivation of the M1 gene abolished the induction of viral enzyme activity in VV-infected cells. Each of the three mutant viruses replicated to levels comparable to the wild-type (WT) virus in BSC40 (monkey), growing A549 (human lung carcinoma) cells, and serum-starved A549 cells, indicating that a functional M1 gene was not required for viral replication in tissue culture. In contrast, in vivo studies indicate that the loss of viral ribonucleotide reductase activity leads to a mild attenuation of VV. By the intracranial route of inoculation, approximately 10-fold more of the M1 lambda recombinant than the WT virus was required to produce the average lethal dose for 50% of the population of injected mice.

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@article{Child1990InsertionalIO, title={Insertional inactivation of the large subunit of ribonucleotide reductase encoded by vaccinia virus is associated with reduced virulence in vivo.}, author={Stephanie J. Child and George Palumbo and R. Mark L. Buller and Dennis E. Hruby}, journal={Virology}, year={1990}, volume={174 2}, pages={625-9} }