Mechanism of Tet(O)-mediated tetracycline resistance

被引:84
作者
Connell, SR
Trieber, CA
Dinos, GP
Einfeldt, E
Taylor, DE
Nierhaus, KH
机构
[1] Max Planck Inst Mol Genet, D-14195 Berlin, Germany
[2] Univ Alberta, Dept Med Microbiol & Immunol, Edmonton, AB T6G 2H7, Canada
[3] Univ Alberta, Dept Biol Sci, Edmonton, AB T6G 2H7, Canada
[4] Univ Patras, Sch Med, Biochem Lab, GR-26110 Patras, Greece
关键词
antibiotic resistance; protein synthesis; ribosome; Tet(O); tetracycline;
D O I
10.1093/emboj/cdg093
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Tet(O) is an elongation factor-like protein which confers resistance to the protein synthesis inhibitor tetracycline by promoting the release of the drug from its inhibitory site on the ribosome. Here we investigated the interaction of Tet(O) with the elongating ribosome and show, using dimethyl sulfate (DMS) probing and binding assays, that it interacts preferentially with the post-translocational ribosome. Furthermore, using an XTP-dependent mutant of Tet(O), we demonstrated that Tet(O) induces conformational rearrangements within the ribosome which can be detected by EF-Tu, and manifested as a stimulation in the GTPase activity of this elongation factor. As such, these conformational changes probably involve the ribosomal GTPase-associated center and, accordingly, Tet(O) alters the DMS modification pattern of the L11 region. Additionally, tetracycline binding is associated with an E-a of 58 kJ/mol. These results suggest a model where both Tet(O) and tetracycline induce a conformational change in functionally opposite directions and the Tet(O)-induced conformation persists after it has left the ribosome; this prevents rebinding of the drug while allowing productive A-site occupation by a ternary complex in the presence of tetracycline.
引用
收藏
页码:945 / 953
页数:9
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