Two-dimensional polyacrylamide gel electrophoresis separation of low molecular weight nuclear RNA

Abstract

Two-dimensional electrophoresis successively on 10%, pH 7.2, and 12%, pH 3.3, polyacrylamide gels was used to separate nuclear and whole cell 4–8S RNA fractions from Novikoff hepatoma ascites cells into individual RNA species. With this method, analytical studies were possible on two new species of RNA referred to as 4.2S RNA and 4.5S RNAII, respectively. The 4.2S RNA has the nucleotide composition AMP, 18.0; UMP, 23.9; GMP, 27.9; and CMP 26.7. This RNA contains 2 ψMP residues and an alkali stable dinucleotide. The 4.5S RNAII was distinctly separated from 4.5S RNAI and 4.5S RNAIII and has the nucleotide composition AMP, 20.6%; UMP, 23.7%; GMP, 30.0%; and CMP, 25.7%. It has no ψMP or 2′-0-methylated nucleotides. In addition, several other spots were separated from the major RNA species.