In vivo and in vitro characterization of protein interactions with the dyad G-box of the Arabidopsis Adh gene.
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'-CCACGTGG-3') is associated with light-induced expression of RbcS and chalcone synthase. The same sequence is also present in the 5'-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.
McKendree
W L
WL
Department of Botany, University of Florida, Gainesville 32611.
Paul
A L
AL
DeLisle
A J
AJ
Ferl
R J
RJ
D000426:Alcohol Dehydrogenase / Q000235:genetics*; D001483:Base Sequence; D002457:Cell Extracts; D002478:Cells, Cultured; D004247:DNA / Q000378:metabolism*; D017343:Genes, Plant / Q000235:genetics*; D008969:Molecular Sequence Data; D010940:Plant Proteins / Q000378:metabolism*; D010944:Plants / Q000201:enzymology* / Q000235:genetics; D012045:Regulatory Sequences, Nucleic Acid / Q000235:genetics
D002457:Cell Extracts; D010940:Plant Proteins; D004247:DNA; D000426:Alcohol Dehydrogenase