Processing of adenovirus RNA before release from isolated nuclei

Nuclei isolated from cultured human cells (KB) infected with adenovirus type-2 were used to study events occurring during the adenosine triphosphate-dependent release of viral RNA. When cells were labeled with [(3)H]uridine for 50 min beginning 18 hr after infection, most nuclear viral messenger RNA was in the form of high molecular weight precursors with sedimentation coefficients greater than 28 S. When nuclei were incubated in the presence of ATP and an ATP-generating system, labeled RNA the size of viral mRNA (10-29 S) was released. Cleavage of viral RNA precursor molecules of high molecular weight occurred during incubation of nuclei. The change in size of viral RNA did not require exogenous ATP and occurred before release of RNA from nuclei. This RNA cleavage seems comparable to processing of mRNA in vivo, for discrete viral species were produced. Cleavage was completed after 10 min of incubation but was not the rate-limiting step in the release reaction, which required about 20 min for completion.