THE PLANT CELL, Vol 2, Issue 3 207-214, Copyright © 1990 by American Society of Plant Biologists
In Vivo and in Vitro Characterization of Protein Interactions with the Dyad G-Box of the Arabidopsis Adh Gene
W. L. McKendree, A. L. Paul, A. J. DeLisle and R. J. Ferl
Departments of Botany and Vegetable Crops, 1253 Fifield Hall, University of Florida, Gainesville, Florida 32611
Expression of the alcohol dehydrogenase (Adh) and ribulose-1,5-bisphosphate
carboxylase small subunit (RbcS) genes of higher plants is
cell-type-specific and environmentally inducible. However, the tissues in
which these two genes are expressed, their modes of induction, and their
protein functions are quite distinct. Adh is expressed in non-green tissue,
induced by anaerobiosis, and repressed in leaves. RbcS is only expressed in
green tissue. An 8-base pair G-box element (5[prime]-CCACGTGG-3[prime]) is
associated with light-induced expression of RbcS and chalcone synthase. The
same sequence is also present in the 5[prime]-flanking region of
Arabidopsis thaliana Adh, and this sequence is associated with a
trans-acting factor in vivo. We report here that in vitro Adh G-box binding
activity is present in crude whole cell extracts of both cell culture and
leaves of Arabidopsis. The authenticity of in vitro Adh G-box binding is
supported by in vivo and in vitro dimethylsulfate footprinting. A clear in
vivo Adh G-box footprint occurs in cell cultures, but comparable in vivo
binding to the Adh G-box does not occur in leaves. Therefore, there does
not appear to be a direct correlation between the presence of the G-box
factor in a tissue and its binding to the Adh G-box.
