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The effect of tRNA binding on the structure of 5 S RNA in Escherichia coli. A chemical modification study.

Göringer, H. Ulrich and Bertram, S. and Wagner, R. :
The effect of tRNA binding on the structure of 5 S RNA in Escherichia coli. A chemical modification study.
In: The Journal of biological chemistry, 259 (1) pp. 491-6. ISSN 0021-9258
[Article] , (1984)

Abstract

The structure of 5 S RNA within the 70 S ribosome from Escherichia coli was studied using the chemical reagent kethoxal (alpha-keto-beta-ethoxybutyraldehyde) to modify accessible guanosines. The modification pattern of 5 S RNA from free 70 S ribosomes was compared with that of poly(U) programmed ribosomes where tRNA had been bound to both the A- and P-sites. Binding to the ribosomal A-site was achieved enzymatically using the elongation factor Tu and GTP in the presence of deacylated tRNA which blocks the ribosomal P-site. Modified guanosines were identified after partial RNase T1 hydrolysis and separation of the hydrolysis products on sequencing gels. Binding of tRNA to the ribosome leads to a strong protection of 5 S RNA guanosine G-41 and to some degree G-44 from kethoxal modification. The limited RNase T1 hydrolysis pattern provides evidence for the existence of a 5 S RNA conformation different from the known 5 S RNA A- and B-forms which are characterized by their gel electrophoretic mobility. The importance of 5 S RNA for the binding of tRNA to the ribosome is discussed.

Item Type: Article
Erschienen: 1984
Creators: Göringer, H. Ulrich and Bertram, S. and Wagner, R.
Title: The effect of tRNA binding on the structure of 5 S RNA in Escherichia coli. A chemical modification study.
Language: English
Abstract:

The structure of 5 S RNA within the 70 S ribosome from Escherichia coli was studied using the chemical reagent kethoxal (alpha-keto-beta-ethoxybutyraldehyde) to modify accessible guanosines. The modification pattern of 5 S RNA from free 70 S ribosomes was compared with that of poly(U) programmed ribosomes where tRNA had been bound to both the A- and P-sites. Binding to the ribosomal A-site was achieved enzymatically using the elongation factor Tu and GTP in the presence of deacylated tRNA which blocks the ribosomal P-site. Modified guanosines were identified after partial RNase T1 hydrolysis and separation of the hydrolysis products on sequencing gels. Binding of tRNA to the ribosome leads to a strong protection of 5 S RNA guanosine G-41 and to some degree G-44 from kethoxal modification. The limited RNase T1 hydrolysis pattern provides evidence for the existence of a 5 S RNA conformation different from the known 5 S RNA A- and B-forms which are characterized by their gel electrophoretic mobility. The importance of 5 S RNA for the binding of tRNA to the ribosome is discussed.

Journal or Publication Title: The Journal of biological chemistry
Volume: 259
Number: 1
Divisions: 10 Department of Biology > Postranscriptional Gene Regulation and RNA Therapeutics
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10 Department of Biology
Date Deposited: 07 Nov 2011 09:11
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