PUTATIVE 15 S PRECURSOR OF GLOBIN MESSENGER-RNA CONTAINS A POLY-(A) SEQUENCE

[3H]Uridine or [3H]adenosine pulse-labelled nuclear RNA was isolated from chicken immature red blood cells and separated on denaturing formamide sucrose gradients. RNA of each gradient fraction was hybridized with unlabelled globin DNA complementary to mRNA (cDNA) and subsequently digested by RNAase A and RNAase T1. The experiments revealed two RNA species with globin coding sequences sedimenting at 9 S and approx. 15 S, the latter probably representing a precursor of 9 S globin mRNA. A poly(A) sequence was demonstrated in this RNA by two different approaches. Nuclear RNA pulse-labelled with [3H]uridine was fractionated by chromatography on poly(U)-Sepharose. Part of the 15 S precursor was found in the poly(A)-containing RNA. In the second approach 15 S RNA pulse-labelled with [3H]adenosine was hybridized with globin cDNA, incubated with RNAase A and RNAase T1 and subjected to chromatography on hydroxyapatite. The hybrids were isolated and after separation of the strands degraded with DNAase I, RNAase A and RNAase T1. By this procedure poly(A) sequences of approximately 100 nucleotides could be isolated from the 15 S RNA with globin coding sequences. The poly(A) sequence was completely degraded by RNAase T2. © 1978.