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. 1985 Jan;82(1):43-7.
doi: 10.1073/pnas.82.1.43.

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

R J Bram, R D Kornberg

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

R J Bram et al. Proc Natl Acad Sci U S A. 1985 Jan.

Abstract

A binding activity specific for the upstream activating sequence of the GAL1-GAL10 promoter of Saccharomyces cerevisiae has been 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 two promoters are closely homologous, with a core consensus sequence of C-G-CG-TG-C-A-A-C-A-G-T-G-C-T-C-C-G-A-A- GC-G-A-T. A synthetic oligonucleotide with such a sequence competes with the upstream activating sequence in the binding reaction.

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