RNA-metabolism in Streptomyces hydrogenans. Effect of 20beta-hydroxysteroid-dehydrogenase inducing dienediol on RNA-content and RNA-profile.

For the isolation of RNA and polysomes, logarithmically growing cells were disrupted by grinding with kieselgur. Other methods failed to produce undergraded m-RNA as shown by 10-50% sucrose gradient-centrifugation and electrophoresis in 2% mixed agarose-acrylamide gel columns. RNA was extracted by a modified phenol method or in presence of DEP. After the application of dienediol, the amount of acid precipitable RNA decreased by 20 to 50% in 20 min. Incorportation of precursors into RNA was much slower immediately after induction but increased during the next two hours. Three hours after induction there was no difference in the RNA-content of induced and control cultures. Degradation of stable as well as of unstable RNA was observed. Simultaneous addition of the inducer and rifamycin inhibited the production of 20beta-STDH suggesting the synthesis of special m-RNA. The effects of some other antibiotics have also been studied. The existence of a specific m-RNA in the induced system was established from differences in the 3H/14C quotients of gel fractions in the region between light r-RNA and t-RNA.