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J Bacteriol, May 1998, p. 2682-2688, Vol. 180, No. 10
0021-9193/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Negative Regulation of IS2 Transposition by the Cyclic AMP (cAMP)-cAMP Receptor Protein Complex

Shiau-Ting Hu,* Hsuan-Chen Wang, Guang-Sheng Lei, and Shao-Hung Wang

Department of Microbiology and Graduate Institute of Microbiology and Immunology, National Yang-Ming University, Taipei, Taiwan, Republic of China

Received 16 October 1997/Accepted 17 March 1998

Three sequences similar to that of the consensus binding sequence of the cyclic AMP (cAMP)-cAMP receptor protein (CRP) complex were found in the major IS2 promoter region. Experiments were performed to determine whether the cAMP-CRP complex plays a role in the regulation of IS2 transposition. In the gel retardation assay, the cAMP-CRP complex was found to be able to bind the major IS2 promoter. A DNA footprinting assay confirmed that the cAMP-CRP complex binds to the sequences mentioned above. With an IS2 promoter-luciferase gene fusion construct, the cAMP-CRP complex was shown to inhibit transcription from the major IS2 promoter. IS2 was found to transpose at a frequency approximately 200-fold higher in an Escherichia coli host defective for CRP or adenyl cyclase than in a wild-type host. These results suggest that the cAMP-CRP complex is a negative regulator of IS2 transposition.

* Corresponding author. Mailing address: Department of Microbiology, National Yang-Ming University, Taipei, Taiwan. Phone: 011-886-2-2826-7107. Fax: 011-886-2-2821-2880. E-mail: tingnahu{at}mailsrv.ym.edu.tw.

J Bacteriol, May 1998, p. 2682-2688, Vol. 180, No. 10
0021-9193/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


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Copyright © 1998 by the American Society for Microbiology. All rights reserved.