THE PLANT CELL, Vol 4, Issue 4 485-496, Copyright © 1992 by American Society of Plant Biologists

RESEARCH ARTICLES

Sequences Flanking the Hexameric G-Box Core CACGTG Affect the Specificity of Protein Binding

M. E. Williams, R. Foster and N. H. Chua
Laboratory for Plant Molecular Biology, The Rockefeller University, 1230 York Avenue, New York, New York 10021-6399

The CACGTG G-box motif is a highly conserved DNA sequence that has been identified in the 5[prime] upstream region of plant genes exhibiting regulation by a variety of environmental signals and physiological cues. Gel mobility shift assays using a panel of G-box oligonucleotides differing in their flanking sequences identified two types of binding activity (A and B) in a cauliflower nuclear extract. Competition gel retardation assays demonstrated that the two types of binding activity were distinct. Type A binding activity interacted with oligonucleotides designated as class I elements, whereas type B binding activity interacted strongly with class II elements and weakly with class I elements. A third class of elements, null elements, did not exhibit any detectable binding under our assay conditions. Gel retardation analysis of nonpalindromic hybrid G-box oligonucleotides indicated that hybrid elements of the same class exhibited binding affinity commensurate with the affinity of the weaker element, hybrid class I/II elements exhibited only type B binding, and hybrid class I/null and class II/null elements did not show any detectable binding activity. These binding activities can be explained by the affinity of bZip G-box binding homo- or heterodimer subunits for G-box half sites. These experiments led to a set of classification rules that can predict the binding activity of all reported plant G-box motifs containing the consensus hexameric core. Tissue- and/or development-specific expression of genes containing G-box motifs may be regulated by the affinity of G-box proteins for the different classes of G-box elements.

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