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Journal of Bacteriology, December 2008, p. 7925-7931, Vol. 190, No. 24
0021-9193/08/$08.00+0     doi:10.1128/JB.00512-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Flexibility of Vibrio cholerae ToxT in Transcription Activation of Genes Having Altered Promoter Spacing{triangledown}

Michelle Bellair and Jeffrey H. Withey*

Department of Immunology and Microbiology, Wayne State University School of Medicine, Detroit, Michigan 48201

Received 15 April 2008/ Accepted 29 September 2008

Cholera, a severe diarrheal disease, is caused by ingestion of the gram-negative bacterium Vibrio cholerae. Expression of V. cholerae virulence factors is highly regulated at the transcriptional and posttranscriptional levels by a complex network of proteins and small noncoding RNAs. The direct activator of transcription of most V. cholerae virulence genes is the ToxT protein. ToxT binds to a 13-bp sequence, the toxbox, located upstream of genes in its regulon. However, the organization of toxboxes relative to each other and to the core promoter elements at different genes varies dramatically. At different ToxT-activated genes a single toxbox may be necessary and sufficient for full activation, or pairs of toxboxes organized as either inverted or direct repeats may be required for full activation. Although all toxboxes are located at positions consistent with a class I promoter architecture, the locations of toxboxes relative to the transcription start site also vary from gene to gene. To further assess the ability of ToxT to activate transcription from different configurations relative to the core promoter elements, we constructed promoter-lacZ fusions having altered spacing both between toxbox pairs and between the promoter-proximal toxbox and the –35 box at five different ToxT-activated promoters. Our results suggest that that ToxT has remarkable flexibility in its positioning as a transcription activator and that different interactions between ToxT and RNA polymerase occur during transcription activation of promoters having different toxbox configurations.


* Corresponding author. Mailing address: Department of Immunology and Microbiology, Wayne State University School of Medicine, 540 E. Canfield, Detroit, MI 48201. Phone: (313) 577-1316. Fax: (313) 577-1155. E-mail: jwithey{at}med.wayne.edu

{triangledown} Published ahead of print on 10 October 2008.


Journal of Bacteriology, December 2008, p. 7925-7931, Vol. 190, No. 24
0021-9193/08/$08.00+0     doi:10.1128/JB.00512-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.