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THE PLANT CELL, Vol 4, Issue 10 1295-1307, Copyright © 1992 by American Society of Plant Biologists
A Maize Protein Associated with the G-Box Binding Complex Has Homology to Brain Regulatory Proteins
N. C. de Vetten, G. Lu and R. J. Ferl
Program in Plant Molecular and Cellular Biology, Horticultural Sciences Department, Institute for Food and Agricultural Sciences, University of Florida, Gainesville, Florida 32611
The G-box element is a moderately conserved component of the promoter of
many inducible genes, including the alcohol dehydrogenase genes of
Arabidopsis and maize. We used monoclonal antibodies generated against
partially purified G-box binding factor (GBF) activity to characterize
maize proteins that are part of the DNA binding complex. Antibodies
interacted with partially purified maize GBF complexes to produce a slower
migrating complex in the gel retardation assay. Immunoprecipitation
experiments suggested that the protein recognized by the antibody is not a
DNA binding protein in and of itself, but rather is associated with the DNA
binding complex. These monoclonal antibodies were used to isolate cDNA
clones encoding a protein that we have designated GF14. Maize GF14 contains
a region resembling a leucine zipper and acidic carboxy and amino termini,
of which the latter can form an amphipathic [alpha]-helix similar to known
transcriptional activators such as VP16 and GAL4. Protein gel blot analysis
of cell culture extract showed that a single, major protein of
approximately 30 kD is recognized by anti-GF14; the protein is also present
predominantly in the kernel and root. The deduced amino acid sequence of
maize GF14 is more than 80% identical to Arabidopsis GF14 and Oenothera
PHP-O, and is more than 60% identical to a class of mammalian brain
proteins described as both protein kinase C inhibitors and activators of
tyrosine and tryptophan hydroxylases. GF14 is found in a variety of
monocotyledons and dicotyledons, gymnosperms, and yeast. This suggests a
deep evolutionary conservation of a potential regulatory protein associated
with a core sequence found in the promoter region of many genes.
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