Specific protein binding to far upstream activating sequences in polymerase II promoters.

Abstract

A binding activity specific for the upstream activating sequence of the GAL1-GAL10 promoter of Saccharomyces cerevisiae was purified 220-fold on the basis of a nitrocellulose filter-binding assay. The binding activity is enriched in a nuclear preparation and is likely to be the GAL4 gene product. DNase I-protection mapping patterns reveal binding to two 30-base-pair regions at the boundaries of the sequence. A nearly identical mapping pattern is obtained with the coordinately regulated GAL7 promoter. The four 30-base-pair regions of binding in the 2 promoters are closely homologous, with a core consensus sequence which is given. A synthetic oligonucleotide with such a sequence completes with the upstream activating sequence in the binding reaction.