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

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 b...

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Bibliographic Details
Published in:The Journal of biological chemistry 2003-01, Vol.278 (2), p.983-990
Main Authors: Lee, Sung-Jae, Engelmann, Afra, Horlacher, Reinhold, Qu, Qiuhao, Vierke, Gudrun, Hebbeln, Carina, Thomm, Michael, Boos, Winfried
Format: Article
Language:English
Subjects:
Amino Acid Sequence
Archaeal Proteins - isolation & purification
Archaeal Proteins - physiology
ATP-Binding Cassette Transporters - genetics
Base Sequence
Binding Sites
Maltose - genetics
Maltose - metabolism
Molecular Sequence Data
Multigene Family
Operon
Repressor Proteins - isolation & purification
Repressor Proteins - physiology
Thermococcus - chemistry
Thermococcus - genetics
Transcription, Genetic
Trehalose - metabolism
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Summary: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.
ISSN:0021-9258
1083-351X
DOI:10.1074/jbc.M210236200