Double-stranded RNA and the enzymology of interferon action

Extracts from interferon-treated, not virus-infected Ehrlich ascites tumor cells differ in various biochemical characteristics from extracts of control cells. We studied three enzymes whose level is enhanced in cells upon treatment with IF and which are causing some of the differences. (2'-5')(A)n synthetase, an enzyme converting ATP into a series of (2'-5') linked oligoadenylates ((2'-5')(An)) in the presence of dsRNA was purified to homogeneity and characterized. The second enzyme, RNase L, a latent endonuclease, which can be activated by (2'-5')(A)n to cleave single-stranded RNAs, was purified several hundredfold. The activation of this enzyme is reversible and is lost upon removal of (2'-5')(A)n. The activation is not accompanied by a large change in shape of conformation of the enzyme. The third enzyme is a protein kinase which if activated by dsRNA can phosphorylate the peptide chain initiation factor eIF-2 and a protein designated P1 of 67,000 daltons. This enzyme was purified several thousandfold. The most highly purified preparation consists of three proteins with P1 as the most abundant component.