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Journal of Bacteriology, November 2002, p. 5971-5978, Vol. 184, No. 21
0021-9193/02/$04.00+0     DOI: 10.1128/JB.184.21.5971-5978.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Environmental Response and Autoregulation of Clostridium difficile TxeR, a Sigma Factor for Toxin Gene Expression

Nagraj Mani,1,{dagger} Dena Lyras,2 Lisa Barroso,3,{ddagger} Pauline Howarth,2 Tracy Wilkins,3 Julian I. Rood,2 Abraham L. Sonenshein,1* and Bruno Dupuy4

Department of Molecular Biology and Microbiology, Tufts University School of Medicine, Boston, Massachusetts 02111,1 Department of Microbiology, Monash University, Clayton, Victoria, Australia,2 Fralin Biotechnology Center, Virginia Tech, Blacksburg, Virginia 24061,3 Unité de Génétique Moléculaire Bactérienne, Institut Pasteur, Paris, France4

Received 23 May 2002/ Accepted 7 August 2002

TxeR, a sigma factor that directs Clostridium difficile RNA polymerase to recognize the promoters of two major toxin genes, was shown to stimulate its own synthesis. Whether expressed in C. difficile, Clostridium perfringens, or Escherichia coli, TxeR stimulated transcription of fusions of the txeR promoter region to reporter genes. As is the case for the tox genes, txeR expression was responsive to the cellular growth phase and the constituents of the medium. That is, the level of expression in broth culture was low during the exponential growth phase, but rapidly increased as cells approached the stationary phase. In the presence of excess glucose, expression from the txeR promoter was repressed. The results support a model for toxin gene expression in which synthesis of TxeR is induced by specific environmental signals. The increased level of TxeR then permits high-level expression of the toxin genes. The study of txeR gene regulation in C. difficile was made possible by introduction of a mobilizable, replicative plasmid via conjugation with E. coli.

* Corresponding author. Mailing address: Department of Molecular Biology and Microbiology, Tufts University School of Medicine, 136 Harrison Ave., Boston, MA 02111. Phone: (617) 636-6761. Fax: (617) 636-0337. E-mail: linc.sonenshein{at}tufts.edu.

{dagger} Present address: Vertex Pharmaceuticals, Inc., Cambridge, MA 02139.

{ddagger} Present address: TechLab, Inc., Blacksburg, VA 24060.

Journal of Bacteriology, November 2002, p. 5971-5978, Vol. 184, No. 21
0021-9193/02/$04.00+0     DOI: 10.1128/JB.184.21.5971-5978.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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