Transit of tRNA through the Escherichia coli ribosome: cross-linking of the 3' end of tRNA to ribosomal proteins at the P and E sites

FEBS Letters
S V KirillovRobert Zimmermann

Abstract

Photoreactive derivatives of yeast tRNA(Phe) containing 2-azidoadenosine at their 3' termini were used to trace the movement of tRNA across the 50S subunit during its transit from the P site to the E site of the 70S ribosome. When bound to the P site of poly(U)-programmed ribosomes, deacylated tRNA(Phe), Phe-tRNA(Phe) and N-acetyl-Phe-tRNA(Phe) probes labeled protein L27 and two main sites within domain V of the 23S RNA. In contrast, deacylated tRNA(Phe) bound to the E site in the presence of poly(U) labeled protein L33 and a single site within domain V of the 23S rRNA. In the absence of poly(U), the deacylated tRNA(Phe) probe also labeled protein L1. Cross-linking experiments with vacant 70S ribosomes revealed that deacylated tRNA enters the P site through the E site, progressively labeling proteins L1, L33 and, finally, L27. In the course of this process, tRNA passes through the intermediate P/E binding state. These findings suggest that the transit of tRNA from the P site to the E site involves the same interactions, but in reverse order. Moreover, our results indicate that the final release of deacylated tRNA from the ribosome is mediated by the F site, for which protein L1 serves as a marker. The results also show that the...Continue Reading

References

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Citations

Mar 12, 2004·International Journal of Antimicrobial Agents·Bülent Bozdogan, Peter C Appelbaum
May 7, 2009·Nucleic Acids Research·Shinichiro ShojiKurt Fredrick
Jun 15, 2007·Applied and Environmental Microbiology·Clemente I MonteroRobert M Kelly
Dec 29, 2005·Proceedings of the National Academy of Sciences of the United States of America·Letizia BrandiClaudio O Gualerzi
Sep 2, 2004·Protein Science : a Publication of the Protein Society·Hongfei WangShigeyuki Yokoyama
Dec 7, 2005·Journal of Molecular Biology·Natalia NevskayaStanislav Nikonov
Sep 4, 2007·Trends in Biochemical Sciences·Andrei Korostelev, Harry F Noller
Sep 9, 2006·Science·Maria SelmerV Ramakrishnan
Nov 11, 2021·Microbiology and Molecular Biology Reviews : MMBR·Madhan R TirumalaiGeorge E Fox

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