Mapping of the second tetracycline binding site on the ribosomal small subunit of E.coli.
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.
Anokhina
Maria M
MM
Department of Chemistry, Moscow State University, 119992 Moscow, Russian Federation.
Barta
Andrea
A
Nierhaus
Knud H
KH
Spiridonova
Vera A
VA
Kopylov
Alexei M
AM
D000900:Anti-Bacterial Agents / Q000737:chemistry / Q000378:metabolism*; D001665:Binding Sites; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D008958:Models, Molecular; D011500:Protein Synthesis Inhibitors / Q000737:chemistry / Q000378:metabolism*; D012336:RNA, Ribosomal, 16S / Q000737:chemistry / Q000378:metabolism*; D012270:Ribosomes / Q000737:chemistry / Q000378:metabolism*; D013752:Tetracycline / Q000737:chemistry / Q000378:metabolism*
D000900:Anti-Bacterial Agents; D011500:Protein Synthesis Inhibitors; D012336:RNA, Ribosomal, 16S; D013752:Tetracycline