Abstract
Abstract The primary sequence of nucleotides has been defined for 4.5 S RNAi, which is one type of low molecular weight nuclear RNA of Novikoff hepatoma ascites cells. This RNA is the first unique nuclear species of mammalian RNA to be sequenced. It is specifically localized to the extranucleolar portion of the nucleus and is not a precursor of tRNA. Highly labeled 4.5 S RNAi was obtained from cells incubated with [32P]orthophosphate in vitro. The RNA was purified by successive gel electrophoresis and chromatography on DEAE-Sephadex. The primary sequence of nucleotides of this RNA shows that it has a purine-rich 5' end and a pyrimidine-rich 3' end: [see PDF for sequence].
Highlights
4.5 S RNA1, which is one type of low molecular weight nuclear RNA of Novikoff hepatoma ascites cells
Labeled 4.5 S RNA1 was obtained from cells incubated with [azP]orthophosphate in vitro
The primary sequence of nucleotides of this RNA shows that it has a purine-rich 5’ end and a pyrimidine-rich 3’ end: pppG-G-U-C-G-A-G-A-G-G-A-U-G-G-C
Summary
Preparation of Radioactive RNA-Preparation of cells and incubation conditions were the same as those reported earlier (4). The 4 to 8 S RNA was separated by sucrose density gradient centrifugation and total 4.5 S. RNA was isolated by preparative polyacrylamide gel electrophoresis (3, 15). The 4.5 S RNA1 was purified by chromatography on DEAE-Sephadex A-50 columns at pH 5.1 (3). The fractions containing 4.5 S RNA1 were pooled and precipitated with 2 volumes of ethanol containing 2% potassium acetate. The precipitated RNA was collected on Millipore filters and dissolved in small volumes of distilled water. Carrier 4.5 S RNA was added to make a total of 0.2 mg and the RNA was precipitated by adding 2 volumes of ethanol containing 2y0 potassium acetate at -20” overnight.
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