TAILIEUCHUNG - Báo cáo khoa học: Structural origins for selectivity and specificity in an engineered bacterial repressor–inducer pair

The bacterial tetracycline transcription regulation system mediated by the tetracycline repressor (TetR) is widely used to study gene expression in prokaryotes and eukaryotes. To study multiple genes in parallel, a triple mutant TetR(K 64 L 135 I 138 ) has been engineered that is selectively induced by the synthetic tetracycline derivative 4-de-dimethylamino-anhydrotetracy-cline (4-ddma-atc) and no longer by tetracycline, the inducer of wild-type TetR. In the present study | Structural origins for selectivity and specificity in an engineered bacterial repressor-inducer pair 1 1- 2 3 3iA ir 2 Michael A. Klieber Oliver Scholz Susanne Lochner Peter Gmeiner Wolfgang Hillen and Yves A. Muller1 1 Lehrstuhlfur Biotechnik Department of Biology Friedrich-Alexander University Erlangen-Nuremberg Germany 2 Lehrstuhl fur Mikrobiologie Department of Biology Friedrich-Alexander University Erlangen-Nuremberg Germany 3 Lehrstuhlfur Pharmazeutische Chemie Department of Chemistry and Pharmacy Friedrich-Alexander University Erlangen-Nuremberg Germany Keywords altered inducer selectivity altered inducer specificity bacterialtranscription regulation crystal structures tetracycline repressor Correspondence Y. A. Muller Lehrstuhlfur Biotechnik Department of Biology Friedrich-Alexander University Erlangen-Nuremberg Im IZMP Henkestrasse 91 D-91052 Erlangen Germany Fax 49 0 9131 8523080 Tel 49 0 9131 8523081 E-mail ymuller@ Present address Department of Biochemistry University of Zurich Switzerland Database Structuraldata are available from the Protein Data Bank under the accession numbers 3FK6 for TetR K64L135I138 alone and 3FK7 for the 4-ddma-atc complex Received 10 March 2009 revised 9 July 2009 accepted 31 July 2009 doi The bacterial tetracycline transcription regulation system mediated by the tetracycline repressor TetR is widely used to study gene expression in prokaryotes and eukaryotes. To study multiple genes in parallel a triple mutant TetR K64L135I138 has been engineered that is selectively induced by the synthetic tetracycline derivative 4-de-dimethylamino-anhydrotetracy-cline 4-ddma-atc and no longer by tetracycline the inducer of wild-type TetR. In the present study we report the crystal structure of TetR K64L135I138 in the absence and in complex with 4-ddma-atc at resolutions of A. Analysis of the structures in light of the available binding data and previously reported TetR complexes .

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