GcvA and Lrp are both necessary for activation of the gcv operon. The upstream GcvA-binding sites 3 and 2 were separated from the Lrp-binding region and the rest of the gcv control region. Moving these sites by 1 or 2 helical turns of DNA further from the gcv promoter reduces, but does not eliminate, either GcvA-mediated activation or repression of a gcvT::lacZ gene fusion. However, moving these sites by 1.5 or 2.5 helical turns of DNA results in a GcvA-mediated super-repression of the operon. This repression is dependent on Lrp and is partially dependent on GcvR. Lrp bound to the gcv control region induces a bend in the DNA. Based on these results, a model for gcv regulation is presented in which Lrp plays a primarily structural role, by bending the DNA and GcvA functions as the activator protein.
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http://dx.doi.org/10.1099/00221287-144-10-2865 | DOI Listing |
Microbiology (Reading)
November 2000
Department of Microbiology, The University of Iowa, Iowa City, IA 52242, USA1.
GcvA binds to three sites in the gcvTHP control region, from base -34 to -69 (site 1), from base -214 to -241 (site 2) and from base -242 to -271 (site 3). Previous results suggested that sites 3 and 2 are required for both GcvA-dependent activation and repression of a gcvT::lacZ fusion. However, the results were less clear as to the role of site 1.
View Article and Find Full Text PDFMicrobiology (Reading)
August 1999
The GcvA protein both activates and represses the gcv operon and negatively regulates its own transcription. GcvA binds to three sites in the gcv control region and to one site in the gcvA control region; each of these binding sites contains the conserved 5 bp DNA sequence 5'-CTAAT-3'. This report describes the role this DNA sequence plays in autoregulation and expression of gcvA.
View Article and Find Full Text PDFMicrobiology (Reading)
October 1998
GcvA and Lrp are both necessary for activation of the gcv operon. The upstream GcvA-binding sites 3 and 2 were separated from the Lrp-binding region and the rest of the gcv control region. Moving these sites by 1 or 2 helical turns of DNA further from the gcv promoter reduces, but does not eliminate, either GcvA-mediated activation or repression of a gcvT::lacZ gene fusion.
View Article and Find Full Text PDFMicrobiology (Reading)
May 1998
Department of Microbiology, University of Iowa, Iowa City, IA 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.
View Article and Find Full Text PDFJ Bacteriol
September 1995
Department of Microbiology, University of Iowa, Iowa City 52242, USA.
The GcvA protein is a LysR family regulatory protein necessary for both activation and repression of the Escherichia coli glycine cleavage enzyme operon (gcv) and negative regulation of gcvA. Gel shift assays indicated that overexpressed GcvA in crude extracts is capable of binding specifically to DNA containing the gcv and gcvA control regions. DNase I footprint analysis of the gcvA control region revealed one region of GcvA-mediated protection overlapping the transcription initiation site and extending from -28 to +20.
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