Journal of Bacteriology, September 2003, p. 5158-5165, Vol. 185, No. 17
0021-9193/03/$08.00+0 DOI: 10.1128/JB.185.17.5158-5165.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.
Determination of the InvE Binding Site Required for Expression of IpaB of the Shigella sonnei Virulence Plasmid: Involvement of a ParB BoxA-Like Sequence
Takayuki Taniya,1,2 Jiro Mitobe,1 Shu-ichi Nakayama,1 Qi Mingshan,3 Kenji Okuda,2 and Haruo Watanabe1*
Department of Bacteriology, National Institute of Infectious Diseases, Toyama 1-23-1, Shinjuku-ku, Tokyo 162-8640,1
Department of Bacteriology, Yokohama City University School of Medicine, Fukuura 3 to 9, Kanazawa-ku, Yokohama 236-0004, Japan,2
Department of Oral Biology, School of Dental Medicine, Buffalo, New York 14214-30923
Received 20 March 2003/
Accepted 16 June 2003
The InvE protein positively regulates the expression of virulence genes ipaBCD in Shigella sonnei. The InvE has significant homology with ParB of plasmid P1, which is known as a plasmid partitioning factor with DNA binding ability. Although the DNA binding activity of InvE has been predicted, it is not known whether the DNA binding activity is necessary for type III secretion system-associated gene expression. In this study, we determined the transcription start site of the icsB-ipaBCD operon (ipa operon) and constructed a series of deletions of the icsB promoter region in the Escherichia coli K-12 background. The deletion study revealed that an 86-bp region upstream of the icsB transcription start site was essential for expression of the ipa operon, where the ParB binding motif (ParB BoxA-like sequence) was observed. Purified glutathione S-transferase-InvE fusion protein bound directly to the -93 to -54 region (designating the icsB transcription start site as nucleotide +1) containing the ParB BoxA-like sequence. These results indicated that InvE bound directly to the promoter region.
* Corresponding author. Mailing address: Department of Bacteriology, National Institute of Infectious Diseases, Toyama 1-23-1, Shinjuku-ku, Tokyo 162-8640, Japan. Phone: (81 3) 5285 1111, ext. 2201. Fax: (81 3) 5285 1171. E-mail: haruwata{at}nih.go.jp.
Journal of Bacteriology, September 2003, p. 5158-5165, Vol. 185, No. 17
0021-9193/03/$08.00+0 DOI: 10.1128/JB.185.17.5158-5165.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.
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Copyright © 2003 by the American Society for Microbiology. All rights reserved.