The behaviour of heterogeneous nuclear ribonucleic acid in partially and completely denaturing conditions

Abstract

It is shown that the heterogeneous nuclear RNA (HnRNA) synthesized in the presence of actinomycin and at low and high temperatures sediments in low-ionic-strength sucrose gradients between the rRNA components, similar to the unmethylated RNA synthesized under ‘step-down’ conditions. If the ionic strength is increased then the HnRNA sediments more rapidly than 28S rRNA, with a large proportion about the 45S precursor rRNA position. Initially this was thought to be due to aggregation of the HnRNA; however, centrifugation and electrophoresis in completely denaturing conditions suggest that the molecular weight of this species of RNA is very large The experiments reveal that HnRNA is conformationally unstable relative to the nucleolar RNA and that the slower sedimentation rate of HnRNA in 5mm-EDTA–Tris base–sucrose gradients reflects the greater expansion of the HnRNA relative to the nucleolar RNA. The implications of this finding are discussed.