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Journal of Bacteriology, December 1999, p. 7558-7565, Vol. 181, No. 24
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Regulation of Sigma 54-Dependent Transcription by Core Promoter Sequences: Role of -12 Region Nucleotides

Lei Wang, Yuli Guo, and Jay D. Gralla*

Department of Chemistry and Biochemistry and Molecular Biology Institute, University of California, Los Angeles, California 90095

Received 29 July 1999/Accepted 29 September 1999

The tetranucleotide core recognition sequence (TTGC) of the sigma 54 promoter -12 recognition element was altered by random substitution. The resulting promoter mutants were characterized in vivo and in vitro. Deregulated promoters were identified, implying that this core element can mediate the response to enhancer-binding proteins. These promoters had in common a substitution at position -12 (consensus C), indicating its importance for keeping basal transcription in check. In another screen, nonfunctional promoters were identified. Their analysis indicated that positions -13 (consensus G) and -15 (consensus T) are important to maintain minimal promoter function. In vitro studies showed that the -13 and -15 positions contribute to closed-complex formation, whereas the -12 position has a stronger effect on recognition of the fork junction intermediate created during open-complex formation. Overall the data indicate that the -12 region core contains specific subsequences that direct the diverse RNA polymerase interactions required both to produce RNA and to restrict this RNA synthesis in the absence of activation.

* Corresponding author. Mailing address: Department of Chemistry and Biochemistry and Molecular Biology Institute, University of California, Los Angeles, CA 90095. Phone: (310) 825-1620. Fax: (310) 267-2302. E-mail: gralla{at}mbi.ucla.edu.

Journal of Bacteriology, December 1999, p. 7558-7565, Vol. 181, No. 24
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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