Localization of the Decoding Region on the 30S Escherichia Coli Ribosomal Subunit by Affinity Immunoelectron Microscopy

Overview

Journal Proc Natl Acad Sci U S A

Specialty Science

Date 1979 Mar 1

PMID 375223

Citations 14

Authors

M Boublik

J OFENGAND

Affiliations

Soon will be listed here.

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.

Citing Articles

Arrangement of the subunits in the ribosome of Escherichia coli: demonstration by immunoelectron microscopy.

Kastner B, Stoffler-Meilicke M, Stoffler G Proc Natl Acad Sci U S A. 1981; 78(11):6652-6.

PMID: 7031657 PMC: 349107. DOI: 10.1073/pnas.78.11.6652.

Localization of 3' ends of 5S and 23S rRNAs in reconstituted subunits of Escherichia coli ribosomes.

Stoffler-Meilicke M, Stoffler G, Odom O, Zinn A, Kramer G, Hardesty B Proc Natl Acad Sci U S A. 1981; 78(9):5538-42.

PMID: 7029538 PMC: 348781. DOI: 10.1073/pnas.78.9.5538.

Localization of the 3' end of 16S rRNA in Escherichia coli 30S ribosomal subunits by immuno electron microscopy.

Luhrmann R, Stoffler-Meilicke M, Stoffler G Mol Gen Genet. 1981; 182(3):369-76.

PMID: 7029211 DOI: 10.1007/BF00293924.

Nascent polypeptide chains emerge from the exit domain of the large ribosomal subunit: immune mapping of the nascent chain.

Bernabeu C, Lake J Proc Natl Acad Sci U S A. 1982; 79(10):3111-5.

PMID: 6808502 PMC: 346363. DOI: 10.1073/pnas.79.10.3111.

Ribosomal protein S4 is an internal protein: Localization by immunoelectron microscopy on protein-deficient subribosomal particles.

Winkelmann D, Kahan L, Lake J Proc Natl Acad Sci U S A. 1982; 79(17):5184-8.

PMID: 6752941 PMC: 346859. DOI: 10.1073/pnas.79.17.5184.

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