Binding interaction between Tet(M) and the ribosome: requirements for binding.
Tet(M) protein interacts with the protein biosynthesis machinery to render this process resistant to tetracycline by a mechanism which involves release of the antibiotic from the ribosome in a reaction dependent on GTP hydrolysis. To clarify this resistance mechanism further, the interaction of Tet(M) with the ribosome has been examined by using a gel filtration assay with radioactively labelled Tet(M) protein. The presence of GTP and 5'-guanylyl imido diphosphate, but not GDP, promoted Tet(M)-ribosome complex formation. Furthermore, thiostrepton, which inhibits the activities of elongation factor G (EF-G) and EF-Tu by binding to the ribosome, blocks stable Tet(M)-ribosome complex formation. Direct competition experiments show that Tet(M) and EF-G bind to overlapping sites on the ribosome.
Department of Microbiology, Duke University Medical Center, Durham, North Carolina 27710, USA.
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D000900:Anti-Bacterial Agents / Q000494:pharmacology; D001426:Bacterial Proteins / Q000187:drug effects / Q000378:metabolism*; D001665:Binding Sites; D004926:Escherichia coli / Q000187:drug effects / Q000378:metabolism*; D006160:Guanosine Triphosphate / Q000378:metabolism; D006165:Guanylyl Imidodiphosphate / Q000378:metabolism; D020653:Peptide Elongation Factor G; D010445:Peptide Elongation Factors / Q000378:metabolism; D012270:Ribosomes / Q000187:drug effects / Q000378:metabolism*; D013753:Tetracycline Resistance / Q000502:physiology*; D013883:Thiostrepton / Q000494:pharmacology
D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; D020653:Peptide Elongation Factor G; D010445:Peptide Elongation Factors; C083138:Tet M resistance protein, Bacteria; D006165:Guanylyl Imidodiphosphate; D006160:Guanosine Triphosphate; D013883:Thiostrepton