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. 1975 Nov;72(11):4332–4336. doi: 10.1073/pnas.72.11.4332

Identification of a region in 23S rRNA located at the peptidyl transferase center.

N Sonenberg, M Wilchek, A Zamir
PMCID: PMC388715  PMID: 1105580

Abstract

A photolyzable derivative of dipeptidyl-tRNA, p-azido-N-tBoc-Phe-[3H]Phe-tRNA, bound reversibly to 70S ribosomes in the presence of poly(U), becomes, when irradiated, covalently attached to components of the 50S ribosomal subunit. Most of the reaction occurs within the 23S RNA, while ribosomal proteins are only weakly labeled. Reversible binding as well as the covalent reaction are reduced in the absence of poly(U) or in the presence of several antibiotics specific for the 50S ribosomal subunit. There are apparently few sites (or perhaps a single site) of reaction on the 23S RNA that are exclusively located within the 2000 nucleotides from the 3 terminus of the molecule. Part of the 23S RNA within this region must therefore be closely associated with the peptidyl transferase center of the ribosome.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Allet B., Spahr P. F. Binding sites of ribosomal proteins on two specific fragments derived from Escherichia coli 50 S ribosomes. Eur J Biochem. 1971 Mar 11;19(2):250–255. doi: 10.1111/j.1432-1033.1971.tb01311.x. [DOI] [PubMed] [Google Scholar]
  2. Barta A., Kuechler E., Branlant C., Sri Widada J., Krol A., Ebel J. P. Photoaffinity labelling of 23 S RNA at the donor-site of the Escherichia coli ribosome. FEBS Lett. 1975 Aug 1;56(1):170–174. doi: 10.1016/0014-5793(75)80134-7. [DOI] [PubMed] [Google Scholar]
  3. Bispink L., Matthaei H. Photoaffinity labeling of 23 S rRNA in Escherichia coli ribosomes with poly(U)-coded ethyl 2-diazomalonyl-Phe-tRNA. FEBS Lett. 1973 Dec 1;37(2):291–294. doi: 10.1016/0014-5793(73)80480-6. [DOI] [PubMed] [Google Scholar]
  4. Cahn F., Schachter E. M., Rich A. Polypeptide synthesis with ribonuclease-digested ribosomes. Biochim Biophys Acta. 1970;209(2):512–520. doi: 10.1016/0005-2787(70)90748-3. [DOI] [PubMed] [Google Scholar]
  5. Cooperman B. S., Jaynes E. N., Brunswick D. J., Luddy M. A. Photoincorporation of puromycin and N-(ethyl-2-diazomalonyl)puromycin into Escherichia coli ribosomes. Proc Natl Acad Sci U S A. 1975 Aug;72(8):2974–2978. doi: 10.1073/pnas.72.8.2974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Czernilofsky A. P., Kuechler E. Affinity label for the tRNA binding site on the Escherichia coli ribosome. Biochim Biophys Acta. 1972 Jul 31;272(4):667–671. doi: 10.1016/0005-2787(72)90526-6. [DOI] [PubMed] [Google Scholar]
  7. Dahlberg A. E., Dingman C. W., Peacock A. C. Electrophoretic characterization of bacterial polyribosomes in agarose-acrylamide composite gels. J Mol Biol. 1969 Apr 14;41(1):139–147. doi: 10.1016/0022-2836(69)90131-4. [DOI] [PubMed] [Google Scholar]
  8. Ginzburg I., Miskin R., Zamir A. N-ethyl maleimide as a probe for the study of functional sites and conformations of 30 S ribosomal subunits. J Mol Biol. 1973 Sep 25;79(3):481–494. doi: 10.1016/0022-2836(73)90400-2. [DOI] [PubMed] [Google Scholar]
  9. Girshovich A. S., Bochkareva E. S., Kramarov V. M., Ovchinnikov YuA E. coli 30 S and 50 S ribosomal subparticle components in the localization region of the tRNA acceptor terminus. FEBS Lett. 1974 Sep 1;45(1):213–217. doi: 10.1016/0014-5793(74)80847-1. [DOI] [PubMed] [Google Scholar]
  10. Greenwell P., Harris R. J., Symons R. H. Affinity labelling of 23-S ribosomal RNA in the active centre of Escherichia coli peptidyl transferase. Eur J Biochem. 1974 Dec 2;49(3):539–544. doi: 10.1111/j.1432-1033.1974.tb03858.x. [DOI] [PubMed] [Google Scholar]
  11. Hartman K. A., Amaya J., Schachter E. M. Structure of RNA in ribosomes. Science. 1970 Oct 9;170(3954):171–173. doi: 10.1126/science.170.3954.171. [DOI] [PubMed] [Google Scholar]
  12. Held W. A., Ballou B., Mizushima S., Nomura M. Assembly mapping of 30 S ribosomal proteins from Escherichia coli. Further studies. J Biol Chem. 1974 May 25;249(10):3103–3111. [PubMed] [Google Scholar]
  13. Held W. A., Gette W. R., Nomura M. Role of 16S ribosomal ribonucleic acid and the 30S ribosomal protein S12 in the initiation of natural messenger ribonucleic acid translation. Biochemistry. 1974 May 7;13(10):2115–2122. doi: 10.1021/bi00707a019. [DOI] [PubMed] [Google Scholar]
  14. Hsiung N., Cantor C. R. A new simpler photoaffinity analogue of peptidyl tRNA. Nucleic Acids Res. 1974 Dec;1(12):1753–1762. doi: 10.1093/nar/1.12.1753. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Hsiung N., Reines S. A., Cantor C. R. Investigation of the ribosomal peptidyl transferase center using a photoaffinity label. J Mol Biol. 1974 Oct 5;88(4):841–855. doi: 10.1016/0022-2836(74)90403-3. [DOI] [PubMed] [Google Scholar]
  16. Katz G., Dudock B. S. Oligonucleotide sequences in wheat germ phenylalanine transfer ribonucleic acid. J Biol Chem. 1969 Jun 10;244(11):3062–3068. [PubMed] [Google Scholar]
  17. Lapidot Y., Elat D., Rappoport S., de Groot N. Peptidyl-tRNA. IX. The use of o-nitrophenylsulfenyl group as N-protecting group in the synthesis of peptidyl-tRNA. Biochem Biophys Res Commun. 1970 Feb 20;38(4):559–563. doi: 10.1016/0006-291x(70)90617-0. [DOI] [PubMed] [Google Scholar]
  18. Miskin R., Zamir A. Enhancement of peptidyl transferase activity by antibiotics acting on the 50 S ribosomal subunit. J Mol Biol. 1974 Jul 25;87(1):121–134. doi: 10.1016/0022-2836(74)90564-6. [DOI] [PubMed] [Google Scholar]
  19. Noller H. F., Chaires J. B. Functional modification of 16S ribosomal RNA by kethoxal. Proc Natl Acad Sci U S A. 1972 Nov;69(11):3115–3118. doi: 10.1073/pnas.69.11.3115. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Oen H., Pellegrini M., Cantor C. R. Peptidyl transferase inhibitors alter the covalent reaction of BrAcPhe-tRNA with the E. coli ribosome. FEBS Lett. 1974 Sep 1;45(1):218–222. doi: 10.1016/0014-5793(74)80848-3. [DOI] [PubMed] [Google Scholar]
  21. Oen H., Pellegrini M., Eilat D., Cantor C. R. Identification of 50S proteins at the peptidyl-tRNA binding site of Escherichia coli ribosomes. Proc Natl Acad Sci U S A. 1973 Oct;70(10):2799–2803. doi: 10.1073/pnas.70.10.2799. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Peacock A. C., Dingman C. W. Molecular weight estimation and separation of ribonucleic acid by electrophoresis in agarose-acrylamide composite gels. Biochemistry. 1968 Feb;7(2):668–674. doi: 10.1021/bi00842a023. [DOI] [PubMed] [Google Scholar]
  23. Pestka S. Studies on the formation of transfer ribonucleic acid-ribosome complexes. IV. A new assay for codon recognition and interaction of transfer ribonucleic acid with 50 S subunits. J Biol Chem. 1968 Aug 10;243(15):4038–4044. [PubMed] [Google Scholar]
  24. SARIN P. S., ZAMECNIK P. C. ON THE STABILITY OF AMINOACYL-S-RNA TO NUCLEOPHILIC CATALYSIS. Biochim Biophys Acta. 1964 Dec 16;91:653–655. doi: 10.1016/0926-6550(64)90018-0. [DOI] [PubMed] [Google Scholar]
  25. Saha B. K. Cleavage of 23-S RNA in 50-S ribosomes into a 5'-terminal and a 3'-terminal fragment by pancreatic ribonuclease. Biochim Biophys Acta. 1974 Jul 11;353(3):292–300. doi: 10.1016/0005-2787(74)90022-7. [DOI] [PubMed] [Google Scholar]
  26. Schwyzer R., Caviezel M. p-Azido-L-phenylalanine: a photo-affinity 'probe' related to tyrosine. Helv Chim Acta. 1971;54(5):1395–1400. doi: 10.1002/hlca.19710540521. [DOI] [PubMed] [Google Scholar]
  27. Sonenberg N., Wilchek M., Zamir A. Mapping of Escherichia coli ribosomal components involved in peptidyl transferase activity. Proc Natl Acad Sci U S A. 1973 May;70(5):1423–1426. doi: 10.1073/pnas.70.5.1423. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Sopori M., Pellegrini M., Lengyel P., Cantor C. R. Affinity labeling of Escherichia coli ribosomal proteins with an analog of the natural initiator tRNA. Biochemistry. 1974 Dec 17;13(26):5432–5439. doi: 10.1021/bi00723a030. [DOI] [PubMed] [Google Scholar]
  29. Spierer P., Zimmerman R. A., Mackie G. A. RNA-protein interactions in the ribosome. Binding of 50-S-subunit proteins to 5' and 3' terminal segments of the 23-S RNA. Eur J Biochem. 1975 Apr 1;52(3):459–468. doi: 10.1111/j.1432-1033.1975.tb04014.x. [DOI] [PubMed] [Google Scholar]
  30. Yukioka M., Hatayama T., Morisawa S. Affinity labeling of the ribonucleic acid component adjacent to the peptidyl recognition center of peptidyl transferase in Escherichia coli ribosomes. Biochim Biophys Acta. 1975 May 1;390(2):192–208. doi: 10.1016/0005-2787(75)90341-x. [DOI] [PubMed] [Google Scholar]

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