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Journal of Bacteriology, February 2006, p. 1316-1331, Vol. 188, No. 4
0021-9193/06/$08.00+0     doi:10.1128/JB.188.4.1316-1331.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

The Transcriptional Antiterminator RfaH Represses Biofilm Formation in Escherichia coli

Christophe Beloin,2,{dagger} Kai Michaelis,1,{dagger} Karin Lindner,1 Paolo Landini,3 Jörg Hacker,1 Jean-Marc Ghigo,2 and Ulrich Dobrindt1*

Institut für Molekulare Infektionsbiologie, Bayerische Julius-Maximilians-Universität Würzburg, Röntgenring 11, 97070 Würzburg, Germany,1 Groupe de Génétique des Biofilms, Institut Pasteur, URA CNRS 2172, 25 rue du Docteur Roux, 75724 Paris Cedex 15, France,2 Swiss Federal Institute of Environmental Technology (EAWAG), Überlandstrasse 133, 8600 Dübendorf, Switzerland3

Received 27 May 2005/ Accepted 23 November 2005

We investigated the influence of regulatory and pathogenicity island-associated factors (Hha, RpoS, LuxS, EvgA, RfaH, and tRNA5Leu) on biofilm formation by uropathogenic Escherichia coli (UPEC) strain 536. Only inactivation of rfaH, which encodes a transcriptional antiterminator, resulted in increased initial adhesion and biofilm formation by E. coli 536. rfaH inactivation in nonpathogenic E. coli K-12 isolate MG1655 resulted in the same phenotype. Transcriptome analysis of wild-type strain 536 and an rfaH mutant of this strain revealed that deletion of rfaH correlated with increased expression of flu orthologs. flu encodes antigen 43 (Ag43), which mediates autoaggregation and biofilm formation. We confirmed that deletion of rfaH leads to increased levels of flu and flu-like transcripts in E. coli K-12 and UPEC. Supporting the hypothesis that RfaH represses biofilm formation through reduction of the Ag43 level, the increased-biofilm phenotype of E. coli MG1655rfaH was reversed upon inactivation of flu. Deletion of the two flu orthologs, however, did not modify the behavior of mutant 536rfaH. Our results demonstrate that the strong initial adhesion and biofilm formation capacities of strain MG1655rfaH are mediated by both increased steady-state production of Ag43 and likely increased Ag43 presentation due to null rfaH-dependent lipopolysaccharide depletion. Although the roles of rfaH in the biofilm phenotype are different in UPEC strain 536 and K-12 strain MG1655, this study shows that RfaH, in addition to affecting the expression of bacterial virulence factors, also negatively controls expression and surface presentation of Ag43 and possibly another Ag43-independent factor(s) that mediates cell-cell interactions and biofilm formation.

* Corresponding author. Mailing address: Institut für Molekulare Infektionsbiologie, Röntgenring 11, D-97070 Würzburg, Germany. Phone: 49 (0)931 312155. Fax: 49 (0)931 312578. E-mail: ulrich.dobrindt{at}mail.uni-wuerzburg.de.

{dagger} C.B. and K.M. contributed equally to this work.

Journal of Bacteriology, February 2006, p. 1316-1331, Vol. 188, No. 4
0021-9193/06/$08.00+0     doi:10.1128/JB.188.4.1316-1331.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



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