Infectivity titration, sedimentation analysis in sucrose gradients and electron microscopy have been used to study virus maturation following the reversal of the inhibition of vaccinia virus growth by rifampicin.

Electron-dense inclusions containing tubular structures develop in the cytoplasm of infected BHK 21/C 13 cells maintained in rifampicin, but the formation of immature and mature virus particles is prevented. The removal of rifampicin is followed by a rise of the virus infectivity. Spicule-covered membranes appear at the periphery of the inclusions and both immature and mature virus particles are seen. A proportion of the DNA synthesized in the presence of rifampicin is incorporated into particles and becomes resistant to deoxyribonuclease I. If protein synthesis is inhibited, spicule-covered membranes and immature particles appear but no mature particles are seen; the virus infectivity does not increase and the DNA remains susceptible to deoxyribonuclease I. It is suggested that rifampicin binds reversibly to a virus-specified protein, thereby preventing the formation of immature virus particles. Possible effects of rifampicin on the subsequent stages of virus maturation are discussed.


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