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Molecular Biology and Gene Regulation

ARF-B₂: A Protein Complex that Specifically Binds to Part of the Anaerobic Response Element of Maize Adh 1¹

Department of Vegetable Crops, University of Florida, Gainesville, Florida 32611

Crude whole cell extracts from maize (Zea mays L.) suspension cells were examined for DNA binding proteins that specifically interact with a portion of the maize Adh 1 promoter that was previously shown to be in contact with a trans-acting factor in vivo. A 17 base pair, double-stranded oligonucleotide probe was constructed that centered around a strong in vivo dimethylsulfate footprint (B₂) that coincides with part of the anaerobic response element (ARE). Gel retardation assays were used to characterize a major, specific DNA binding protein activity found in the crude extracts. The activity is present in both aerobic and hypoxically treated cultures and has been designated ARF-B₂ (ARE binding factor). ARF-B₂ appears to be a multicomponent complex, with a 54 kilodalton subunit termed ARF-B₂ in primary contact with the target DNA.

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