Isolation of informoferes from rat liver: Effects of α-amanitin and actinomycin D

Abstract

RNP particles carrying rapidly labelled RNA (informoferes) were isolated from rat liver nuclei by extraction with isotonic and 0.3 M salt buffer at pH 8 either with or without ultrasonic treatment. The importance of preliminary extraction of the nuclei with the isotonic salt buffer at a lower pH of 7 or in the presence of 50 mM EDTA is demonstrated. Administration of α-amanitin or of actinomycin D, in doses inhibiting the labelling of the heterogeneous RNA with RNA precursors in the range of 60–85%, reduces the amount of informoferes recovered. The informoferes isolated from treated animals are highly depleted in HnRNA. They still contain, however, low molecular RNA species with a slower turnover than the HnRNA. The polypeptide pattern observed after acrylamide gel electrophoresis of the informofere proteins is qualitatively changed in the treated preparations, whereas the ratio of protein to RNA decreases. In the presence of RNase inhibitor the informoferes are recovered in form of polymer structures. α-Amanitin and actinomycin D significantly reduce the amount of polymers recovered.