The role of ribosomal ribonucleic acid in the structure and function of mammalian brain ribosomes.
In order to resolve the functional role of intact rRNA in polypeptide chain elongation mouse brain ribosomes were treated with dilute pancreatic or T(1) RNAase (ribonuclease). After RNAase treatment, several physical-chemical properties as well as the functional activity of the ribosomes were measured. RNAase treatment resulted in the extensive hydrolysis of both 18S and 28S rRNA; however, the sedimentation properties of mono-ribosomes were unaltered and more than 90% of the relatively low-molecular-weight RNA fragments remained associated with ribosome particles. Analysis of the ability of RNAase-treated ribosomes to participate in cell-free protein synthesis showed that ribosomes with less than 2% intact rRNA retained more than 85% of their activity in polyphenylalanine incorporation. Proof that the incorporation of phenylalanine by ribosomes with hydrolysed rRNA actually represented active translocation was obtained by the effective inhibition of incorporation by diphtheria toxin. In addition, the oligopeptide products of protein synthesis could be identified by BD (benzoylated diethylaminoethyl)-cellulose column chromatography. Analysis of the size distribution of oligopeptides synthesized by normal and RNAase-treated ribosomes showed no significant differences which indicated that there was no change in the proportion of ribosomes engaged in protein synthesis. Thus strong RNA-protein and protein-protein interactions must serve to maintain the functional integrity of ribosomes even when the rRNA is extensively degraded. The ability of the enzyme-treated ribosomes to efficiently incorporate amino acids clearly demonstrated that ;intact' rRNA is not required for protein-synthetic activity.
D000818:Animals; D001921:Brain / Q000378:metabolism*; D002250:Carbon Radioisotopes; D002458:Cell Fractionation; D004167:Diphtheria Toxin; D004591:Electrophoresis, Polyacrylamide Gel; D006358:Hot Temperature; D006868:Hydrolysis; D066298:In Vitro Techniques; D051379:Mice; D008970:Molecular Weight; D010452:Peptide Biosynthesis; D010441:Peptide Chain Elongation, Translational; D010649:Phenylalanine / Q000378:metabolism; D011485:Protein Binding; D014176:Protein Biosynthesis; D012335:RNA, Ribosomal / Q000378:metabolism*; D012260:Ribonucleases; D012270:Ribosomes / Q000378:metabolism* / Q000648:ultrastructure
D002250:Carbon Radioisotopes; D004167:Diphtheria Toxin; D012335:RNA, Ribosomal; D010649:Phenylalanine; D012260:Ribonucleases