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Binding of the Escherichia coli MelR protein to the melAB promoter: orientation of MelR subunits and investigation of MelR–DNA contacts by David C. Grainger; Tamara A. Belyaeva; David J. Lee; Eva I. Hyde; Stephen J. W. Busby is a Biochemistry, Genetics and Molecular Biology article available to read on EtoBox.

What is Binding of the Escherichia coli MelR protein to the melAB promoter: orientation of MelR subunits and investigation of MelR–DNA contacts about?

## Summary The __Escherichia coli__ MelR protein is a melibiose‐triggered transcription factor, belonging to the AraC family, that activates transcription initiation at the __melAB__ promoter. Activation is dependent on the binding of MelR to four 18 bp sites, centred at position −42.5 (site 2′), position −62.5 (site 2), position −100.5 (site 1) and position −120.5 (site 1′) relative to the __melAB__ transcription start point. Activation also depends on the binding of CRP to a single site located between MelR binding site 1 and site 2. All members of the AraC family contain two helix–turn–helix (HTH) motifs that contact two segments of the DNA major groove at target sites on the same DNA face. In this work, we have studied the binding of MelR to different sites at the __melAB__ promoter, focusing on the orientation of binding of the two MelR HTH motifs, and the juxtaposition of the different bound MelR subunits with respect to each other. To do this, MelR was engineered to contain a single cysteine residue adjacent to either one or the other HTH motif. The MelR derivatives were purified, and the cysteine residues were tagged with __p__‐bromoacetamidobenzyl‐EDTA‐Fe, an inorgan

Who reads Binding of the Escherichia coli MelR protein to the melAB promoter: orientation of MelR subunits and investigation of MelR–DNA contacts?

Author

David C. Grainger; Tamara A. Belyaeva; David J. Lee; Eva I. Hyde; Stephen J. W. Busby

Publisher

John Wiley and Sons; Wiley (Blackwell Publishing); Blackwell Publishing Inc.; Wiley (ISSN 0950-382X)

Published

2003

Language

EN

Field

Biochemistry, Genetics and Molecular Biology (Life Sciences)