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

Richard J Bram, R. D. Kornberg

Research output: Contribution to journalArticle

117 Citations (Scopus)

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 region.

Original languageEnglish (US)
Pages (from-to)43-47
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume82
Issue number1
StatePublished - 1985
Externally publishedYes

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Protein Binding
Base Pairing
Collodion
Deoxyribonuclease I
Consensus Sequence
Oligonucleotides
Saccharomyces cerevisiae
Genes

ASJC Scopus subject areas

  • General
  • Genetics

Cite this

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AU - Bram, Richard J

AU - Kornberg, R. D.

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N2 - 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 region.

AB - 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 region.

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