Mapping of the second tetracycline binding site on the ribosomal small subunit of E.coli |
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Authors: | Anokhina Maria M Barta Andrea Nierhaus Knud H Spiridonova Vera A Kopylov Alexei M |
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Affiliation: | Department of Chemistry, Moscow State University, 119992 Moscow, Russian Federation. |
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Abstract: | 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. |
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