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

Nucleic Acids Res. 2004 May 11;32(8):2594-7. doi: 10.1093/nar/gkh583. Print 2004.


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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Anti-Bacterial Agents / chemistry
  • Anti-Bacterial Agents / metabolism*
  • Binding Sites
  • Escherichia coli / genetics
  • Escherichia coli / metabolism*
  • Models, Molecular
  • Protein Synthesis Inhibitors / chemistry
  • Protein Synthesis Inhibitors / metabolism*
  • RNA, Ribosomal, 16S / chemistry
  • RNA, Ribosomal, 16S / metabolism*
  • Ribosomes / chemistry
  • Ribosomes / metabolism*
  • Tetracycline / chemistry
  • Tetracycline / metabolism*


  • Anti-Bacterial Agents
  • Protein Synthesis Inhibitors
  • RNA, Ribosomal, 16S
  • Tetracycline

Associated data

  • PDB/1I97