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Microbiology 144 (1998), 1417-1422
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microbiology, Vol 144, 1417-1422, Copyright © 1998 by Society for General Microbiology

ARTICLES

Spacing and orientation requirements of GcvA-binding sites 3 and 2 and the Lrp-binding region for gcvT::lacZ expression in Escherichia coli

LT Stauffer and GV Stauffer
Department of Microbiology, University of Iowa, Iowa City 52242, USA.

Both GcvA and Lrp are required for normal regulation of the gcv operon. Moving the GcvA-binding sites 3 and 2 and the Lrp-binding region either closer to, or further away from, the gcv promoter by approximately one helical turn of DNA resulted in a less than twofold decrease in glycine- mediated activation or inosine-mediated repression of a gcvT::IacZ fusion. Moving these sites approximately two helical turns of DNA away from the gcv promoter resulted in a further loss of both activation and repression; moving these sites approximately three helical turns of DNA from the gcv promoter resulted in an essentially complete loss of both glycine-mediated activation and inosine-mediated repression. However, when these sites were moved by approximately 1.5 and 2.5 helical turns of DNA away from the gcv promoter, there was a complete loss of both glycine-mediated activation and inosine-mediated repression of the gcvT::IacZ fusion. The flexibility in the absolute distance of the GcvA- and Lrp-binding sites relative to the gcv promoter, but strict orientation dependence of these sites is consistent with a possible protein-protein interaction of either GcvA, Lrp, or both of these proteins with RNA polymerase. Because of the location of these target sites relative to the gcv promoter, it is also likely that DNA looping is required for this mechanism of regulation.


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G. Heil, L. T. Stauffer, and G. V. Stauffer
Glycine binds the transcriptional accessory protein GcvR to disrupt a GcvA/GcvR interaction and allow GcvA-mediated activation of the Escherichia coli gcvTHP operon
Microbiology, July 1, 2002; 148(7): 2203 - 2214.
[Abstract] [Full Text] [PDF]

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L. D. Wonderling, M. L. Urbanowski, and G. V. Stauffer
GcvA binding site 1 in the gcvTHP promoter of Escherichia coli is required for GcvA-mediated repression but not for GcvA-mediated activation
Microbiology, November 1, 2000; 146(11): 2909 - 2918.
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 J. Bacteriol.Home page
A. D. Jourdan and G. V. Stauffer
Mutational Analysis of the Transcriptional Regulator GcvA: Amino Acids Important for Activation, Repression, and DNA Binding
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