scholarly journals Differential stability of 28s and 18s rat liver ribosomal ribonucleic acids

1969 ◽  
Vol 115 (1) ◽  
pp. 91-94 ◽  
Author(s):  
P. V. Venkov ◽  
A. A. Hadjiolov
Keyword(s):  
Rat Liver ◽  
Gel Electrophoresis ◽  
18S Rrna ◽  
Heterogeneous Material ◽  
28S Rrna ◽  
Agar Gel ◽  
Ribonucleic Acids ◽  
Different Temperatures ◽  
Rrna Species ◽  
Agar Gel Electrophoresis

Rat liver ribosomal RNA (rRNA) free from nuclease contaminants was isolated by a modification of the phenol technique. The 28s and 18s rRNA species were separated by preparative agar-gel electrophoresis. The two rRNA species were heated at different temperatures under various conditions and the amount of undegraded rRNA was determined by analytical agar-gel electrophoresis. The 18s rRNA remained unaltered after heating for up to 10min. at 90° in water, acetate buffer, pH5·0, or phosphate buffer, pH7·0. Under similar or milder conditions 28s rRNA was partially degraded, giving rise to a well-delimited 6s peak and a heterogeneous material located in the zone between 28s and 6s. The dependence of degradation of 28s rRNA on the temperature and the ionic strength of the medium was studied. The greatest extent of degradation of 28s rRNA was observed on heating at 90° in water. It is suggested that the instability of rat liver 28s rRNA is due to two factors: the presence of hidden breaks in the polymer chain and a higher susceptibility of some phosphodiester bonds to thermal hydrolysis.

scholarly journals Quantitative analysis of rat liver nucleolar and nucleoplasmic ribosomal ribonucleic acids

1978 ◽  
Vol 171 (2) ◽  
pp. 367-374 ◽  
Author(s):  
M D Dabeva ◽  
K P Dudov ◽  
A A Hadjiolov ◽  
A S Stoykova
Keyword(s):  
Gel Electrophoresis ◽  
Molar Ratio ◽  
Absolute Amount ◽  
Agar Gel ◽  
Ribonucleic Acids ◽  
Nuclear Rna ◽  
Rrna Species ◽  
Detergent Treatment ◽  
Agar Gel Electrophoresis ◽  
Nucleolar Rna

rRNA from detergent-purified nuclei was fractionated quantitatively, by two independent methods, into nucleolar and nucleoplasmic RNA fractions. The two RNA fractions were analysed by urea/agar-gel electrophoresis and the amount of pre-rRNA (precursor of rRNA) and rRNA components was determined. The rRNA constitutes 35% of total nuclear RNA, of which two-thirds are in nucleolar RNA and one-third in nucleoplasmic RNA. The identified pre-rRNA components (45 S, 41 S, 39 S, 36 S, 32 S and 21 S) are confined to the nucleolus and constitute about 70% of its rRNA. The remaining 30% are represented by 28 S and 18 S rRNA, in a molar ratio of 1.4. The bulk of rRNA in nucleoplasmic RNA is represented by 28 S and 18 S rRNA in a molar ratio close to 1.0. Part of the mature rRNA species in nucleoplasmic RNA originate from ribosomes attached to the outer nuclear membrane, which resist detergent treatment. The absolute amount of nuclear pre-rRNA and rRNA components was evaluated. The amount of 32 S and 21 S pre-rRNA (2.9 × 10(4) and 2.5 × 10(4) molecules per nucleus respectively) is 2-3-fold higherthan that of 45 S, 41 S and 36 S pre-rRNA.

scholarly journals Intranuclear maturation pathways of rat liver ribosomal ribonucleic acids

1976 ◽  
Vol 160 (3) ◽  
pp. 495-503 ◽  
Author(s):  
M D Dabeva ◽  
K P Dudov ◽  
A A Hadjiolov ◽  
I Emanuilov ◽  
B N Todorov
Keyword(s):  
Secondary Structure ◽  
Rat Liver ◽  
Mammalian Cells ◽  
18S Rrna ◽  
Electron Microscopic Observation ◽  
28S Rrna ◽  
Electron Microscopic ◽  
Rrna Species ◽  
A Minor

The maturation of pre-rRNA (precursor to rRNA)in liver nuclei is studied by agar/ureagel electrophoresis, kinetics of labelling in vivo with [14C] orotate and electron-microscopic observation of secondary structure of RNA molecules. (1) Processing starts from primary pre-rRNA molecules with average mol. wt. 4.6×10(6)(45S) containing the segments of both 28S and 18S rRNA. These molecules form a heterogeneous peak on electrophoresis. The 28S rRNA segment is homogeneous in its secondary structure. However, the large transcribed spacer segment (presumably at the 5′-end) is heterogeneous in size and secondary structure. A minor early labelled RNA component with mol.wt. about 5.8×10(6) is reproducibly found, but its role as a pre-rRNA species remains to be determined. (2) The following intermediate pre-rRNA species are identified: 3.25×10(6) mol.wt.(41S), a precursor common to both mature rRNA species; 2.60×10(6)(36S) and 2.15×10(6)(32S) precursors to 28S rRNA; 1.05×10(6) (21S) precursor to 18S rRNA. The pre-rRNA molecules in rat liver are identical in size and secondary structure with those observed in other mammalian cells. These results suggest that the endonuclease-cleavage sites along the pre-rRNA chain are identical in all mammalian cells. (3) Labelling kinetics and the simultaneous existence of both 36S and 21S pre-rRNA reveal that processing of primary pre-rRNA in adult rat liver occurs simultaneously by at least two major pathways: (i) 45S → 41S → 32S+21S → 28S+18S rRNA and (ii) 45S → 41S → 36S+18S → 32S → 28S rRNA. The two pathways differ by the temporal sequence of endonuclease attack along the 41 S pre-rRNA chain. A minor fraction (mol.wt.2.9×10(6), 39S) is identified as most likely originating by a direct split of 28S rRNA from 45S pre-rRNA. These results show that in liver considerable flexibility exists in the order of cleavage of pre-rRNA molecules during processing.

scholarly journals Extensive homologies between the methylated nucleotide sequences in several vertebrate ribosomal ribonucleic acids

1978 ◽  
Vol 169 (3) ◽  
pp. 531-542 ◽  
Author(s):  
M S N Khan ◽  
M Salim ◽  
B E H. Maden
Keyword(s):  
Xenopus Laevis ◽  
18S Rrna ◽  
Cultured Cells ◽  
Nucleotide Sequences ◽  
Methyl Groups ◽  
28S Rrna ◽  
Chick Embryo Fibroblast ◽  
Ribonucleic Acids ◽  
Rrna Species ◽  
Complete Digestion

The methylated nucleotide sequences in the rRNA molecules of the following vertebrate cultured cells were compared: human (HeLa); hamster (BHK/C13); mouse (L); chick-embryo fibroblast; Xenopus laevis kidney. In each species the combined 18S, 28S and 5.8S molecules possess approx. 110-115 methyl groups, and the methylated oligonucleotides released after complete digestion of the rRNA by T1 ribonuclease encompass several hundred nucleotides. “Fingerprints” of the three mammalian methyl-labelled 18S rRNA species were qualitatively indistinguishable. “Fingerprints” of digests of 28S rRNA of hamster and mouse L-cells were extremely similar to those of HeLa cells, differing in one and three methylated oligonucleotides respectively. “Fingerprints” of methyl-labelled rRNA from chick and Xenopus strongly resembled those of mammals in most respects, but differed in several oligonucleotides in both 18S and 28S rRNA. At least some of the differences between “fingerprints” appear to be due to single base changes or to the presence or absence of methyl groups at particular points in the primary sequence. The findings strongly suggest that the methylated-nucleotide sequences are at least 95% homologous between the rRNA molecules of the two most distantly related vertebrates compared, man and Xenopus laevis.

POLYETHYLENE GLYCOL ON AGAR-GEL ELECTROPHORESIS

The Lancet ◽  
1974 ◽  
Vol 304 (7892) ◽  
pp. 1321-1322
Author(s):  
W.H. Taylor ◽  
D.J. Etherington
Keyword(s):  
Polyethylene Glycol ◽  
Gel Electrophoresis ◽  
Agar Gel ◽  
Agar Gel Electrophoresis
Sign in / Sign up

Export Citation Format

Share Document