Published online 24 June 2004
Nucleic Acids Research, Vol. 32 No. 11 © Oxford University Press 2004; all rights reserved
Extending the classification of bacterial transcription factors beyond the helix–turn–helix motif as an alternative approach to discover new cis/trans relationships
Centre d'Ingénierie des Protéines, Université de Liège, Institut de Chimie B6a, B-4000, Liège, Belgium, 1 Lehrstuhl für Mikrobiologie, Friedrich-Alexander-Universität Erlangen-Nürnberg, Staudtstrasse 5, 91058 Erlangen, Germany and 2 Department of Molecular Microbiology, John Innes Centre, Colney, Norwich NR4 7UH, UK
* To whom correspondence should be addressed. Tel: +32 4 366 33 77; Fax: +32 4 366 33 64; Email: srigali{at}ulg.ac.be
The authors wish it to be known that, in their opinion, the first two authors should be regarded as joint First Authors
Received May 5, 2004; Revised and Accepted June 8, 2004
Transcription factors (TFs) of bacterial helix–turn–helix superfamilies exhibit different effector-binding domains (EBDs) fused to a DNA-binding domain with a common feature. In a previous study of the GntR superfamily, we demonstrated that classifying members into subfamilies according to the EBD heterogeneity highlighted unsuspected and accurate TF-binding site signatures. In this work, we present how such in silico analysis can provide prediction tools to discover new cis/trans relationships. The TF-binding site consensus of the HutC/GntR subfamily was used to (i) predict target sites within the Streptomyces coelicolor genome, (ii) discover a new HutC/GntR regulon and (iii) discover its specific TF. By scanning the S.coelicolor genome we identified a presumed new HutC regulon that comprises genes of the phosphotransferase system (PTS) specific for the uptake of N-acetylglucosamine (PTSNag). A weight matrix was derived from the compilation of the predicted cis-acting elements upstream of each gene of the presumed regulon. Under the assumption that TFs are often subject to autoregulation, we used this matrix to scan the upstream region of the 24 HutC-like members of S.coelicolor. orf SCO5231 (dasR) was selected as the best candidate according to the high score of a 16 bp sequence identified in its upstream region. Our prediction that DasR regulates the PTSNag regulon was confirmed by in vivo and in vitro experiments. In conclusion, our in silico approach permitted to highlight the specific TF of a regulon out of the 673 orfs annotated as ‘regulatory proteins’ within the genome of S.coelicolor.
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