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J. Biol. Chem., Vol. 278, Issue 2, 983-990, January 10, 2003

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TrmB, a Sugar-specific Transcriptional Regulator of the Trehalose/Maltose ABC Transporter from the Hyperthermophilic Archaeon Thermococcus litoralis^*

Sung-Jae Lee, Afra Engelmann^§, Reinhold Horlacher, Qiuhao Qu, Gudrun Vierke^§, Carina Hebbeln^§, Michael Thomm^§, and Winfried Boos^¶

From the  Department of Biology, University of Konstanz, 78457 Konstanz, Germany, and ^§ Institut für Allgemeine Mikrobiologie, Universität Kiel, 24118 Kiel, Germany

We report the characterization of TrmB, a protein of 38,800 apparent molecular weight, that is involved in the maltose-specific regulation of a gene cluster in Thermococcus litoralis, malE malF malG orf trmB malK, encoding a binding protein-dependent ABC transporter for trehalose and maltose. TrmB binds maltose and trehalose half-maximally at 20 µM and 0.5 mM sugar concentration, respectively. Binding of maltose but not of trehalose showed indications of sigmoidality and quenched the intrinsic tryptophan fluorescence by 15%, indicating a conformational change on maltose binding. TrmB causes a shift in electrophoretic mobility of DNA fragments harboring the promoter and upstream regulatory motif identified by footprinting. Band shifting by TrmB can be prevented by maltose. In vitro transcription assays with purified components from Pyrococcus furiosus have been established to show pmalE promoter-dependent transcription at 80 °C. TrmB specifically inhibits transcription, and this inhibition is counteracted by maltose and trehalose. These data characterize TrmB as a maltose-specific repressor for the trehalose/maltose transport operon of Thermococcus litoralis.

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