Covalent Attachment of a Peptidyl-transfer RNA Analog to the 50S Subunit of Escherichia Coli Ribosomes

Overview

Journal Proc Natl Acad Sci U S A

Specialty Science

Date 1972 Apr 1

PMID 4554533

Citations 6

Authors

M Pellegrini

H Oen

C R Cantor

Affiliations

Soon will be listed here.

Abstract

The peptidyl-tRNA analog, N-bromo-acetyl-Phenylalanyl-tRNA(Phe) has been prepared. Its binding to the 70S ribosome of E. coli is totally dependent upon polyuridylic acid. The analog becomes covalently attached to the 50S particle. It is associated with only one protein fraction after polyacrylamide-gel separation of total 50S proteins. The analog also reacts with 23S ribosomal RNA or a protein that remains tightly bound to this RNA after treatment with LiCl-urea and sodium dodecyl sulfate. The analog can function as a peptidyl-tRNA for at least one peptide transfer, but it then inhibits further chain elongation. This result strongly suggests that this analog becomes covalently bound at the P-site of the ribosome.

Citing Articles

Identification of chloramphenicol-binding protein in Escherichia coli ribosomes by affinity labeling.

Pongs O, Bald R, Erdmann V Proc Natl Acad Sci U S A. 1973; 70(8):2229-33.

PMID: 4599619 PMC: 433707. DOI: 10.1073/pnas.70.8.2229.

Proteins at the tRNA binding sites of Escherichia coli ribosomes.

Czernilofsky A, Collatz E, Stoffler G, Kuechler E Proc Natl Acad Sci U S A. 1974; 71(1):230-4.

PMID: 4589893 PMC: 387971. DOI: 10.1073/pnas.71.1.230.

Identification of 50S proteins at the peptidyl-tRNA binding site of Escherichia coli ribosomes.

Oen H, Pellegrini M, Eilat D, Cantor C Proc Natl Acad Sci U S A. 1973; 70(10):2799-803.

PMID: 4583025 PMC: 427112. DOI: 10.1073/pnas.70.10.2799.

Mapping of Escherichia coli ribosomal components involved in peptidyl transferase activity.

Sonenberg N, Wilchek M, Zamir A Proc Natl Acad Sci U S A. 1973; 70(5):1423-6.

PMID: 4576018 PMC: 433511. DOI: 10.1073/pnas.70.5.1423.

Synthesis of cyclohexylpuromycin and its reaction with N-acetylphenylalanyl-transfer ribonucleic acid on rat liver ribosomes.

Ariatti M, Hawtrey A Biochem J. 1975; 145(2):169-76.

PMID: 1156356 PMC: 1165205. DOI: 10.1042/bj1450169.

References

Staehelin T, Maglott D, MONRO R . On the catalytic center of peptidyl transfer: a part of the 50 S ribosome structure. Cold Spring Harb Symp Quant Biol. 1969; 34:39-48. DOI: 10.1101/sqb.1969.034.01.008. View

Nakamoto T, Kalokofsky D . A possible mechanism for initiation of protein synthesis. Proc Natl Acad Sci U S A. 1966; 55(3):606-13. PMC: 224195. DOI: 10.1073/pnas.55.3.606. View

Lanks K, Sciscenti J, Weinstein I, Cantor C . Studies on rat liver phenylalanyl transfer ribonucleic acid synthetase. I. Purification, stabilization, and complex formation. J Biol Chem. 1971; 246(11):3494-9. View

Maden B, Traut R, MONRO R . Ribosome-catalysed peptidyl transfer: the polyphenylalanine system. J Mol Biol. 1968; 35(2):333-45. DOI: 10.1016/s0022-2836(68)80028-2. View

De Groot N, Lapidot Y, Panet A, Wolman Y . The synthesis of N-acetylphenylalanyl-sRNA. Biochem Biophys Res Commun. 1966; 25(1):17-22. DOI: 10.1016/0006-291x(66)90633-4. View

Chang F, Flaks J . Topography of the Escherichia coli ribosome. II. Preliminary sequence of 50 s subunit protein attack by trypsin and its correlation with functional activities. J Mol Biol. 1971; 61(2):387-400. DOI: 10.1016/0022-2836(71)90388-3. View

Shaw E . Selective chemical modification of proteins. Physiol Rev. 1970; 50(2):244-96. DOI: 10.1152/physrev.1970.50.2.244. View

Nomura M, Erdmann V . Reconstitution of 50S ribosomal subunits from dissociated molecular components. Nature. 1970; 228(5273):744-8. DOI: 10.1038/228744a0. View

Traub P, Nomura M . Structure and function of E. coli ribosomes. V. Reconstitution of functionally active 30S ribosomal particles from RNA and proteins. Proc Natl Acad Sci U S A. 1968; 59(3):777-84. PMC: 224743. DOI: 10.1073/pnas.59.3.777. View

10.

Haenni A, Chapeville F . The behaviour of acetylphenylalanyl soluble ribonucleic acid in polyphenylalanine synthesis. Biochim Biophys Acta. 1966; 114(1):135-48. DOI: 10.1016/0005-2787(66)90261-9. View

11.

Chrambach A, Reisfeld R, Wyckoff M, Zaccari J . A procedure for rapid and sensitive staining of protein fractionated by polyacrylamide gel electrophoresis. Anal Biochem. 1967; 20(1):150-4. DOI: 10.1016/0003-2697(67)90272-2. View

12.

Gilbert W . Polypeptide synthesis in Escherichia coli. I. Ribosomes and the active complex. J Mol Biol. 1963; 6:374-88. DOI: 10.1016/s0022-2836(63)80050-9. View

13.

Gilbert W . Polypeptide synthesis in Escherichia coli. II. The polypeptide chain and S-RNA. J Mol Biol. 1963; 6:389-403. DOI: 10.1016/s0022-2836(63)80051-0. View

14.

Nirenberg M, MATTHAEI J . The dependence of cell-free protein synthesis in E. coli upon naturally occurring or synthetic polyribonucleotides. Proc Natl Acad Sci U S A. 1961; 47:1588-602. PMC: 223178. DOI: 10.1073/pnas.47.10.1588. View

15.

Traut R, Delius H, AHMAD-ZADEH C, Bickle T, Pearson P, TISSIERES A . Ribosomal proteins of E. Coli: stoichiometry and implications for ribosome structure. Cold Spring Harb Symp Quant Biol. 1969; 34:25-38. DOI: 10.1101/sqb.1969.034.01.007. View

16.

Wood W, Berg P . The effect of enzymatically synthesized ribonucleic acid on amino acid incorporation by a soluble protein-ribosome system from Escherichia coli. Proc Natl Acad Sci U S A. 1962; 48:94-104. PMC: 285510. DOI: 10.1073/pnas.48.1.94. View

17.

Nomura M . Bacterial ribosome. Bacteriol Rev. 1970; 34(3):228-77. PMC: 378356. DOI: 10.1128/br.34.3.228-277.1970. View

18.

Schofield P, Zamecnik P . Cupric ion catalysis in hydrolysis of aminoacyl-tRNA. Biochim Biophys Acta. 1968; 155(2):410-6. DOI: 10.1016/0005-2787(68)90185-8. View

19.

MONRO R . Catalysis of peptide bond formation by 50 S ribosomal subunits from Escherichia coli. J Mol Biol. 1967; 26(1):147-51. DOI: 10.1016/0022-2836(67)90271-9. View

20.

Smith I, Yamane T . Spin-labeled nucleic acids. Proc Natl Acad Sci U S A. 1967; 58(3):884-7. PMC: 335719. DOI: 10.1073/pnas.58.3.884. View