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PLANT PHYSIOLOGY , Vol 115, Issue 2 397-407, Copyright © 1997 by American Society of Plant Biologists
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GENE REGULATION AND MOLECULAR GENETICS |
A G-Box-Binding Protein from Soybean Binds to the E1 Auxin-Response Element in the Soybean GH3 Promoter and Contains a Proline-Rich Repression Domain
Z. B. Liu, G. Hagen and T. J. Guilfoyle
Department of Biochemistry, 117 Schweitzer Hall, University of Missouri, Columbia, Missouri 65211
The E1 promoter fragment (-249 to -203) is one of three auxin-response
elements (AuxREs) in the soybean (Glycine max L.) GH3 promoter (Z.-B. Liu,
T. Ulmasov, X. Shi, G. Hagen, T.J. Guilfoyle [1994] Plant Cell 6: 645-657).
Results presented here further characterize and delimit the AuxRE within
the E1 fragment. The E1 fragment functioned as an AuxRE in transgenic
tobacco (Nicotiana tabacum L.) plants, as well as in transfected
protoplasts. The AuxRE within E1 contains a G-box, and this G-box was used
to clone a G-box-binding factor (GBF) from soybean (SGBF-2). This 45-kD GBF
contains an N-terminal proline-rich domain and a C-terminal basic/leucine
zipper DNA-binding domain. Gel-mobility shift assays were used to
characterize the binding specificity of SGBF-2. Antiserum raised against
recombinant SGBF-2 was used to further characterize SGBF-2 and
antigenically related GBFs in soybean nuclear extracts. Co-transfection
assays with effector and reporter plasmids in carrot (Daucus carota L.)
protoplasts indicated that the N-terminal proline-rich domain of SGBF-2
functioned as a repression domain in both basal and auxin-inducible
transcription.
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