SPECIFIC PROTEIN-BINDING TO FAR UPSTREAM ACTIVATING SEQUENCES IN POLYMERASE-II PROMOTERS
SPECIFIC PROTEIN-BINDING TO FAR UPSTREAM ACTIVATING SEQUENCES IN POLYMERASE-II PROMOTERS
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DOI：
10.1073/pnas.82.1.43
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发表时间：
1985-01-01
影响因子：
11.1
通讯作者：
KORNBERG, RD
中科院分区：
文献类型：
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作者：
BRAM, RJ;KORNBERG, RD
基于硝酸纤维素过滤结合测定，将对酿酒酵母的GAL 1-GAL 10启动子的上游激活序列特异的结合活性纯化220倍。结合活性在核制备物中富集，并且可能是GAL 4基因产物。DNA酶I-保护映射模式揭示了在序列的边界处与两个30个碱基对区域的结合。一个几乎相同的映射模式，获得与协调调节GAL 7启动子。2个启动子中的4个30个碱基对的结合区域是紧密同源的，具有给出的核心共有序列。具有这种序列的合成寡核苷酸在结合反应中与上游激活序列一起完成。
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.