Studies on the inhibition by α-amanitin of single-step addition reactions and productive RNA synthesis catalysed by wheat-germ RNA polymerase II

Abstract

The rate of formation of a single phosphodiester bond with UTP substrate, U-A primer, poly[d(A-T)] template and wheat-germ RNA polymerase II is greatly depressed in the presence of alpha-amanitin. Half-maximal inhibition occurs at 0.04 microgram/ml, in close agreement with published values for inhibition of productive RNA synthesis with class II RNA polymerases from higher-plant species. However, a sizeable proportion of U-A-U synthesis is resistant to inhibition by excess alpha-amanitin. In the additional presence of ATP, i.e. under experimental conditions permitting RNA chain elongation, the synthesis of poly[r(A-U)] is arrested after the formation of the first phosphodiester bond. The results support the contention that the main enzymic process disrupted by alpha-amanitin is the translocation step of the transcription complex along the DNA template.