Comparative studies of the primary structures of ribosomal RNAs of several eukaryotic cell lines by the fingerprinting method

Abstract

Comparisons of the primary structures of 18S and 28S ribosomal RNAs of man, rat, mouse and chicken were made by two-dimensional fractionation including electrophoresis at pH 3.5 and homochromatography. All large T1 oligonucleotides were recovered from the different fingerprints and their radioactivity was measured. They were then hydrolysed with pancreatic RNase and the pancreatic products were digested with alkali to determine their base composition and detect modified residues. Finally, residues bearing a modification on the ribose were analysed by hydrolyses with snake venom and spleen phosphodiesterases. For the 18A RNAs 23, 27, 26, 24 oligonucleotides, whose lengths range from 22 to 10 residues, were analyzed respectively for man, rat, mouse and chicken. Among these, 14 are identical in the four species, two at least are common to man, rat, mouse but differ by the presence of A-Cps in chicken spot 4' instead of A-Up in spot 4 and A2-Gp in chicken spot 14 instead of A2-Gp in spot 13. For the 28S RNAs of man, rat, mouse and chicken, 20, 19, 21 and 22 oligonucleotides ranging in length from 27 to 12 residues were analyzed. 11 of them are common to the four species; 4 of them are found in man, rat, mouse and one of these (spot 1) has a corresponding spot in chicken from which it differs only by the existence of A3-Up instead of A2-Up. Another mammalian oligonucleotide (spot 6) differs from its homologous chicken spot (spot 6') bytwo point mutations. The same modified residues as found by Khan and Maden in man, chicken, and xenopus, have been found in rat and mouse. Moreover when these modified residues are common to several species they are found within an identical nucleotide sequence, as can be seen in the case of spots 1, 3, 9, 11 of 18S RNAs and 4, 7, 13 for 28S RNAs. The number of differences observed between the ribosomal RNAs of the four species were compared to the number of differences observed in the same species for several proteins, globins alpha and beta, insulin, cytochrome C and lysozyme.