Photo-Affinity Labeling of tRNA Binding Sites in Macromolecules. I. Linking of the Phenacyl- p -azide of 4-Thiouridine in ( Escherichia coli ) Valyl-tRNA to 16S RNA at the Ribosomal P Site
- 1 October 1974
- journal article
- Published by Proceedings of the National Academy of Sciences in Proceedings of the National Academy of Sciences
- Vol. 71 (10) , 3951-3955
- https://doi.org/10.1073/pnas.71.10.3951
Abstract
The phenacyl-p-azide of 4-thiouridine in (E. coli) tRNA(1) (Val) was prepared for use as a photo-affinity probe of tRNA binding sites on ribosomes. The derivatized tRNA was 90-100% as active as control tRNA for aminoacylation, nonenzymatic binding to the ribosomal P site, elongation factor Tu(EFTu)-dependent binding to the A site, EFTu-GTP-aa-tRNA ternary complex formation, and transfer of valine into polypeptide. Irradiation of p-azidophenacyl-[(3)H]valyl-tRNA bound noncovalently to the ribosomal P site resulted in covalent attachment of 15-20% of the noncovalently bound tRNA to the ribosomes. The linking occurred exclusively to the 16S RNA of the 30S ribosomal subunit, thus suggesting that the region of the ribosome within 9 A of the 4-thiouridine of tRNA, when it is bound in the P site, is solely 16S RNA.Keywords
This publication has 21 references indexed in Scilit:
- Function of Individual 30S Subunit Proteins of Escherichia coli . Effect of Specific Immunoglobulin Fragments (Fab) on Activities of Ribosomal Decoding SitesProceedings of the National Academy of Sciences, 1974
- Identification of a protein at the ribosomal donor-site by affinity labelingBiochemical and Biophysical Research Communications, 1974
- Proteins at the tRNA Binding Sites of Escherichia coli RibosomesProceedings of the National Academy of Sciences, 1974
- Identification of 50S Proteins at the Peptidyl-tRNA Binding Site of Escherichia coli RibosomesProceedings of the National Academy of Sciences, 1973
- Use of Protection of 30S Ribosomal Proteins by tRNA for Functional Mapping of the E. coli RibosomeNature New Biology, 1973
- Functional Modification of 16S Ribosomal RNA by KethoxalProceedings of the National Academy of Sciences, 1972
- Functional heterogeneity of the 30S ribosomal subunit ofEscherichia coliMolecular Genetics and Genomics, 1972
- Interaction of fragmented and cross-linked Escherichia coli valine transfer ribonucleic acid with T u factor-guanosine triphosphate complex.1972
- The function of pseudouridylic acid in transfer ribonucleic acid. Irradiation and cyanoethylation of E. coli valine tRNA fragmentsFEBS Letters, 1971
- Reaction of thiouracil and thiouridine with 2-hydroxy-5-nitrobenzyl bromideBiochimica et Biophysica Acta (BBA) - Nucleic Acids and Protein Synthesis, 1971