PMID:10692154

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Citation

Schreiber, V, Steegborn, C, Clausen, T, Boos, W and Richet, E (2000) A new mechanism for the control of a prokaryotic transcriptional regulator: antagonistic binding of positive and negative effectors. Mol. Microbiol. 35:765-76

Abstract

MalT, the transcriptional activator of the Escherichia coli maltose regulon, self-associates, binds promoter DNA and activates initiation of transcription only in the presence of ATP and maltotriose, the inducer. In vivo studies have revealed that MalT action is negatively controlled by the MalY protein. Using a biochemical approach, we analyse here the mechanism whereby MalY represses MalT activity. We show that MalY inhibits transcription activation by MalT in a purified transcription system. In vitro, a constitutive MalT variant (which is partially active in the absence of maltotriose) is less sensitive than wild-type MalT to repression by MalY, as observed in vivo. We demonstrate that MalY forms a complex with MalT only in the absence of maltotriose and that, conversely, MalY inhibits maltotriose binding by MalT. Together, these results establish that MalY acts directly upon MalT without the help of any factor, and that MalY is a negative effector of MalT competing with the inducer for MalT binding.

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Keywords

Bacterial Proteins/genetics; Bacterial Proteins/isolation & purification; Bacterial Proteins/metabolism; Bacterial Proteins/physiology; Binding, Competitive/drug effects; Cystathionine gamma-Lyase/isolation & purification; Cystathionine gamma-Lyase/metabolism; Cystathionine gamma-Lyase/physiology; DNA-Binding Proteins; Escherichia coli/genetics; Escherichia coli/metabolism; Escherichia coli Proteins; Gene Expression Regulation, Bacterial; Genetic Variation; Maltose/metabolism; Mutation; Nucleotides/metabolism; Nucleotides/pharmacology; Protein Binding/drug effects; Repressor Proteins/metabolism; Repressor Proteins/physiology; Transcription Factors; Transcription, Genetic; Trisaccharides/metabolism

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