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Microbiology 144 (1998), 2865-2872; DOI  10.1099/00221287-144-10-2865
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Roles for GcvA-binding sites 3 and 2 and the Lrp-binding region in gcvT:: lacZ expression in Escherichia coli

Lorraine T. Stauffer and George V. Stauffer1w

Department of Iowa, Iowa City, IA 52242, USA University Of Iowa City, IA 52242, USA

1 Author for correspondence: George V. Stauffer. Tel: + 1 319 33.5 7791. Fax: + 1 319 33.5 9006. e-mail: george-stauffer@uiowa.edu

ABSTRACT

SUMMARY: GcvA and Lrp are both necessary for activation of the gcv operon. The upstream GcvA-binding sites 3 and 2 were separated from the Lrplbinding 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:: lac2 gene fusion. However, moving these sites by 1-5 or 2.5 helical turns of DNA results inm 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 gcw regulation is presented in which Lrp plays a primarily structural role, by bending the DNA and GcvA functions as the activator protein.


Keywords: glycine cleavage, gcvT:: lacz, GcvA, Lrp, DNA bending




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