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  • Title: Specific protein binding to the simian virus 40 enhancer in vitro.
    Author: Wildeman AG, Zenke M, Schatz C, Wintzerith M, Grundström T, Matthes H, Takahashi K, Chambon P.
    Journal: Mol Cell Biol; 1986 Jun; 6(6):2098-105. PubMed ID: 3023918.
    Abstract:
    HeLa cell nuclear extracts and wild-type or mutated simian virus 40 enhancer DNA were used in DNase I footprinting experiments to study the interaction of putative trans-acting factors with the multiple enhancer motifs. We show that these nuclear extracts contain proteins that bind to these motifs. Because point mutations which are detrimental to the activity of a particular enhancer motif in vivo specifically prevent protection of that motif against DNase I digestion in vivo, we suggest that the bound proteins correspond to trans-acting factors involved in enhancement of transcription. Using mutants in which the two domains A and B of the simian virus 40 enhancer are either separated by insertion of DNA fragments or inverted with respect to their natural orientation, we also demonstrate that the trans-acting factors bind independently to the two domains.
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