PMID: 375223Mar 1, 1979Paper

Localization of the decoding region on the 30S Escherichia coli ribosomal subunit by affinity immunoelectron microscopy

Proceedings of the National Academy of Sciences of the United States of America
M Keren-ZurJ Ofengand

Abstract

The decoding region of the Escherichia coli ribosome has been localized by affinity immunoelectron microscopy. Valyl-tRNA1Val, derivatized at the alpha-amino end with the dinitrophenyl group, was bound to the ribosomal P site of 70S ribosomes and crosslinked specifically to 16S RNA by 310- to 325-nm irradiation. Previous work has shown that crosslink occurs between the 5' anticodon base of the tRNA and a pyrimidine in the 3' one-third of the 16S RNA. By reaction of the dinitrophenyl group with antibody, dimers of the 30S tRNA covalent complexes were prepared containing one covalently attached tRNA per 30S subunit. Electron microscopic visualization of the antibody attached to the dinitrophenyl group located the position of the 3' end of the tRNA. Several sites, with a strong preference for the large protrusion or cleft region in the upper one-third of the subunit, were found. The multiplicity of sites is likely due to the freedom of orientation of the 3' end of the tRNA which is approximately 80 A from the site of crosslinking. By extrapolating this distance from each of the antigenic sites, we concluded that the anticodon of tRNA when in the P site is probably located in the cleft region of the 30S subunit.

References

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Citations

Jan 1, 1982·Progress in Biophysics and Molecular Biology·A Liljas
Apr 1, 1992·Biochimie·E N Trifonov
Jan 1, 1986·Bio Systems·S A Gerbi
Feb 1, 1980·Proceedings of the National Academy of Sciences of the United States of America·H M OlsonB S Cooperman
Nov 1, 1981·Proceedings of the National Academy of Sciences of the United States of America·B KastnerG Stöffler
Sep 1, 1981·Proceedings of the National Academy of Sciences of the United States of America·M Stöffler-MeilickeB Hardesty
May 1, 1982·Proceedings of the National Academy of Sciences of the United States of America·C Bernabeu, J A Lake
Sep 1, 1982·Proceedings of the National Academy of Sciences of the United States of America·D A WinkelmannJ A Lake
Jan 1, 1986·Proceedings of the National Academy of Sciences of the United States of America·M I OakesJ A Lake
Sep 1, 1981·Journal of Ultrastructure Research·I E Ivanov, D D Sabatini
Sep 5, 1981·Journal of Molecular Biology·J M Robertson, W Wintermeyer
Jan 1, 1980·Annals of the New York Academy of Sciences·B S Cooperman
Jan 1, 1980·Annals of the New York Academy of Sciences·J OfengandM Boublik
Jan 8, 1999·Journal of Molecular Biology·C MerrymanH F Noller
Jun 13, 1983·FEBS Letters·A S Spirin
Nov 15, 1982·European Journal of Biochemistry·N RiehlB Ehresmann
Sep 1, 1980·Journal of Virology·B I MilavetzJ A Huberman

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