Journal Article

Mapping of the second tetracycline binding site on the ribosomal small subunit of <i>E.coli</i>

Maria M. Anokhina, Andrea Barta, Knud H. Nierhaus, Vera A. Spiridonova and Alexei M. Kopylov

in Nucleic Acids Research

Volume 32, issue 8, pages 2594-2597
Published in print April 2004 | ISSN: 0305-1048
Published online April 2004 | e-ISSN: 1362-4962 | DOI: http://dx.doi.org/10.1093/nar/gkh583
Mapping of the second tetracycline binding site on the ribosomal small subunit of E.coli

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Tetracycline blocks stable binding of aminoacyl‐tRNA to the bacterial ribosomal A‐site. Various tetracycline binding sites have been identified in crystals of the 30S ribosomal small subunit of Thermus thermophilus. Here we describe a direct photo‐ affinity modification of the ribosomal small subunits of Escherichia coli with 7‐[3H]‐tetracycline. To select for specific interactions, an excess of the 30S subunits over tetracycline has been used. Primer extension analysis of the 16S rRNA revealed two sites of the modifications: C936 and C948. Considering available data on tetracycline interactions with the prokaryotic 30S subunits, including the presented data (E.coli), X‐ray data (T.thermophilus) and genetic data (Helicobacter pylori, E.coli), a second high affinity tetracycline binding site is proposed within the 3′‐major domain of the 16S rRNA, in addition to the A‐site related tetracycline binding site.

Journal Article.  3134 words.  Illustrated.

Subjects: Chemistry ; Biochemistry ; Bioinformatics and Computational Biology ; Genetics and Genomics ; Molecular and Cell Biology

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