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Journal of Bacteriology, June 1999, p. 3864-3868, Vol. 181, No. 12
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Activation of Escherichia coli leuV Transcription by FIS

Wilma Ross,1 Julia Salomon,1 Walter M. Holmes,2 and Richard L. Gourse1,*

Department of Bacteriology, University of Wisconsin, Madison, Wisconsin 53706,1 and Department of Microbiology and The Massey Cancer Center, Medical College of Virginia, Virginia Commonwealth University, Richmond, Virginia 232982

Received 10 February 1999/Accepted 16 April 1999

The transcription factor FIS has been implicated in the regulation of several stable RNA promoters, including that for the major tRNALeu species in Escherichia coli, tRNA1Leu. However, no evidence for direct involvement of FIS in tRNA1Leu expression has been reported. We show here that FIS binds to a site upstream of the leuV promoter (centered at -71) and that it directly stimulates leuV transcription in vitro. A mutation in the FIS binding site reduces transcription from a leuV promoter in strains containing FIS but has no effect on transcription in strains lacking FIS, indicating that FIS contributes to leuV expression in vivo. We also find that RNA polymerase forms an unusual heparin-sensitive complex with the leuV promoter, having a downstream protection boundary of ~-7, and that the first two nucleotides of the transcript, GTP and UTP, are required for formation of a heparin-stable complex that extends downstream of the transcription start site. These studies have implications for the regulation of leuV transcription.

* Corresponding author. Mailing address: Dept. of Bacteriology, University of Wisconsin, 1550 Linden Dr., Madison, WI 53706. Phone: (608) 262-9813. Fax: (608) 262-9865. E-mail: rgourse{at}bact.wisc.edu.

Journal of Bacteriology, June 1999, p. 3864-3868, Vol. 181, No. 12
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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Copyright © 1999 by the American Society for Microbiology. All rights reserved.