In order to study interrelationships between the components of the interferon enzyme system and the cyclic AMP system, NIH 3T3 cells were incubated in the presence of theophylline or adrenaline that cause a rise of intracellular cAMP, respectively, through inhibition of phosphodiesterase of cAMP and activation of adenylate cyclase. In doses that caused a transient, 2- to 3-fold elevation of the cAMP level, theophylline and adrenaline elicited about 2.5-fold elevation of 2′,5′-oligoadenylate synthetase (2-5A synthetase) activity. This increase could be prevented by actinomycin D. This suggests that the elevation of the enzyme activity in the cells was due to a transcription-dependent induction process. Theophylline and adrenaline treatment of the cell cultures also led to a 2-to 3-fold fall of the activity of the phosphodiesterase of 2′,5′-oligoadenylate (2′-phosphodiesterase). This effect of adrenaline was prevented by propanolol but not by actinomycin D. In the case of adrenaline, the fall of 2′-phosphodiesterase activity was accompanied by at least 5-fold increase in the enzyme activity which did not occur if actinomycin D was present in the culture. Similarities and differences between these effects and those induced by interferon are discussed. It is concluded that CAMP is an important regulator of the enzyme system of the 2′,5′-oligoadenylate metabolism. 2′,5′-Oligoadenylate, in turn, was found to act on the activity of phosphodiesterase of cyclic AMP. The cAMP phosphodiesterase activity in the NIH 3T3 cell lysates was activated 2- to 2.5-fold at physiological concentrations (10 −9 to 10 −7 m) of both the phosphorylated form of oligoisoadenylate, ppp(5′A2′p) n5′A2′OH, and the dephosphorylated form, HO(5′A2′p) 25′A2′OH. The phosphorylated form of oligoisoadenylate also activated partially purified preparations of cAMP phosphodiesterase. The data obtained in this study allow us to consider cAMP and 2′,5′-oligoadenylate as the key metabolites that may be used in the cells to form a complex, interconnected, multifunctional circuit that involves the interferon enzyme system and the system of cyclic AMP metabolism and governs essential cell functions, as regulation of RNA metabolism and protein synthesis, cell growth and differentiation.