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Journal Article

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


Nierhaus,  Knud H.
Ribosomes, Max Planck Institute for Molecular Genetics, Max Planck Society;

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Anokhina, M. M., Barta, A., Nierhaus, K. H., Spiridonova, V. A., & Kopylov, A. M. (2004). Mapping of the second tetracycline binding site on the ribosomal small subunit of E.coli. Nucleic Acids Research, 32(8), 2594-2597. doi:DOI: 10.1093/nar/gkh583.

Cite as: https://hdl.handle.net/11858/00-001M-0000-0010-8851-A
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.