Synthesis and Assembly of Simian Virus 40 I. Differential Synthesis of Intact Virions and Empty Shells

Abstract

Intact virions and empty shells of simian virus 40 may be rapidly separated from each other and from cell contaminants by a procedure employing a CsCl cushion. This approach permits quantitation of their respective syntheses in infected cells labeled with radioactive amino acids. As much as 5 to 10% of the total acid-precipitable radioactive lysine in infected cell extracts was incorporated into viral particles in a two-hour pulse late in infection. Evidence for multiple origins of empty shells is presented. Some of the empty shells result from breakdown of intact virions. However, empty shells can also form independently of intact virions. First, labeling for periods of 15 min to 2 hr late in the course of infection results in preferential incorporation of 3 H-lysine into empty shells. Secondly, treatment with the deoxyribonucleic acid inhibitor cytosine-β- d -arabinofuranoside late in infection results in a 50% inhibition in the rate of formation of intact virions with minimal reduction in the rate of appearance of empty shells.