Maximal transcriptional activation by the IHF protein of Escherichia coli depends on optimal DNA bending by the activator

Authors

  • Manuel Engelhorn,

    1. Department of Molecular Biology, University of Geneva, 30, Quai Ernest Ansermet, CH-1211 Geneva 4, Switzerland.,
    Search for more papers by this author
  • Johannes Geiselmann

    1. Laboratoire du Contrôle de l'Expression Génique, Université Joseph Fourier, CERMO, 460, rue de la Piscine, BP53X, 38041 Grenoble Cedex 9, France.
    Search for more papers by this author

Johannes Geiselmann E-mail Hans.Geiselmann@ujf-grenoble.fr; Tel. (4) 76 63 56 62; Fax (4) 76 51 43 36.

Abstract

Transcriptional activation in prokaryotes can be mediated by at least two different mechanisms: direct contacts between the activator and RNA polymerase or modulation of the overall geometry of DNA. In the latter case, an activator protein that bends DNA favours contacts between the DNA upstream of the activator binding site and the back of RNA polymerase. The architectural protein integration host factor (IHF) of Escherichia coli bends DNA and activates transcription at several promoters. We have isolated mutants of IHF that maximize transcriptional activation by adjusting the bending angle of the DNA. The amino acid residues of IHF that adjust the bending angle are close to the DNA and probably make electrostatic interactions with the DNA. We show that transcriptional activation is maintained when the IHF binding site is moved further upstream or when its orientation is inverted, and we conclude from these data that direct interactions between IHF and RNA polymerase do not participate in activation. IHF acts merely by bending DNA; weaker bending leading to stronger activation. We propose that wild-type IHF induces too strong a DNA bend (180°) for optimal interactions between DNA upstream of the IHF binding site and the back of RNA polymerase.

Ancillary