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Mapping of the second tetracycline binding site on the ribosomal small subunit of E.coli.

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.

SUBMITTER: Anokhina MM

PROVIDER: S-EPMC419471 | biostudies-literature | 2004

REPOSITORIES: biostudies-literature

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Mapping of the second tetracycline binding site on the ribosomal small subunit of E.coli.

Anokhina Maria M MM Barta Andrea A Nierhaus Knud H KH Spiridonova Vera A VA Kopylov Alexei M AM

Nucleic acids research 20040511 8

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 ...[more]

PMID: 15141029

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