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Published online 7 August 2000. doi:10.1083/jcb.150.3.447
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© The Rockefeller University Press, 0021-9525/2000//447 $5.00
The Journal of Cell Biology, Volume 150, Number 3, , 2000 447-460

Original Article

Visualization of Trna Movements on the Escherichia coli 70s Ribosome during the Elongation Cycle



Rajendra K. Agrawala, Christian M.T. Spahnb,c, Pawel Penczeka, Robert A. Grassuccib, Knud H. Nierhausc, and Joachim Franka,b

a Wadsworth Center, Department of Biomedical Sciences, State University of New York, Albany, New York 12201
b Howard Hughes Medical Institute, Health Research, Incorporated at Wadsworth Center, Albany, New York 12201
c Max-Planck-Institut für Molekulare Genetik, Ihnestrasse 73, D-14195 Berlin, Germany
HHMI, Wadsworth Center, Empire State Plaza, PO Box 509, Albany, NY 12201-0509.518 486-2191518 474-7002

joachim{at}wadsworth.org

Three-dimensional cryomaps have been reconstructed for tRNA–ribosome complexes in pre- and posttranslocational states at 17-Å resolution. The positions of tRNAs in the A and P sites in the pretranslocational complexes and in the P and E sites in the posttranslocational complexes have been determined. Of these, the P-site tRNA position is the same as seen earlier in the initiation-like fMet-tRNAfMet-ribosome complex, where it was visualized with high accuracy. Now, the positions of the A- and E-site tRNAs are determined with similar accuracy. The positions of the CCA end of the tRNAs at the A site are different before and after peptide bond formation. The relative positions of anticodons of P- and E-site tRNAs in the posttranslocational state are such that a codon–anticodon interaction at the E site appears feasible.

Key Words: ribosomes • protein synthesis • tRNA-binding sites • cryoelectron microscopy • elongation cycle

© 2000 The Rockefeller University Press

Abbreviations used in this paper: 3D, three-dimensional; ASL, anticodon stem-loop; cryo-EM, cryoelectron microscopy; EF-G, elongation factor G; EF-Tu, elongation factor Tu; MF-mRNA, 46-nt long mRNA with AUG and UUC codon in its middle; POST, posttranslocational; PRE, pretranslocational.


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