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J. Biol. Chem., Vol. 277, Issue 8, 5866-5874, February 22, 2002
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From the Department of Microbiology, Cornell University, Ithaca,
New York 14853
OccR is a LysR-type transcriptional regulator of
Agrobacterium tumefaciens that positively regulates the
octopine catabolism operon of the Ti plasmid and is also an
autorepressor. Positive control of the occ genes occurs in
response to octopine, a nutrient released from crown gall tumors. OccR
binds to a site upstream of the occQ promoter in the
presence and absence of octopine. Octopine causes prebound OccR to
undergo a conformational change at the DNA binding site that causes
changes in footprint length and DNA bending. To determine the roles of
these conformational changes in transcriptional activation, we isolated
22 OccR mutants that were able to activate the occQ
promoter in the absence of octopine. Thirteen of these mutants
contained single amino acid substitutions, and nine contained two base
pair changes resulting in two amino acid substitutions, which in
most cases acted synergistically. These mutations spanned the entire
length of the protein. Most of these mutant proteins in the absence of
octopine displayed DNA binding and bending properties
characteristic of transcriptionally active OccR-octopine complexes.
Constitutive Mutations of the OccR Regulatory Protein Affect DNA
Bending in Response to Metabolites Released from Plant Tumors*
*
This work was supported by National Research Service Award
GM41892 from the National Institute of General Medical Sciences.The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
To whom correspondence should be addressed: Dept. of Microbiology,
360A Wing Hall, Cornell University, Ithaca, NY 14853. Tel.: 607-255-2413; Fax: 607-255-3904; E-mail: scw2@cornell.edu.
Copyright © 2002 by The American Society for Biochemistry and Molecular Biology, Inc.
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