Reverse two-hybrid and one-hybrid systems to detect dissociation of protein-protein and DNA-protein interactions

Marc Vidal, Rainer K. Brachmann, Ali Fattaey, Ed Harlow, Jef D. Boeke

    Research output: Contribution to journalArticle

    Abstract

    Macromolecular interactions define many biological phenomena. Although genetic methods are available to identify novel protein-protein and DNA- protein interactions, no genetic system has thus far been described to identify molecules or mutations that dissociate known interactions. Herein, we describe genetic systems that detect such events in the yeast Saccharomyces cerevisiae. We have engineered yeast strains in which the interaction of two proteins expressed in the context of the two-hybrid system or the interaction between a DNA-binding protein and its binding site in the context of the one-hybrid system is deleterious to growth. Under these conditions, dissociation of the interaction provides a selective growth advantage, thereby facilitating detection. These methods referred to as the 'reverse two-hybrid system' and 'reverse one-hybrid system' facilitate the study of the structure-function relationships and regulation of protein- protein and DNA-protein interactions. They should also facilitate the selection of dissociator molecules that could be used as therapeutic agents.

    Original languageEnglish (US)
    Pages (from-to)10315-10320
    Number of pages6
    JournalProceedings of the National Academy of Sciences of the United States of America
    Volume93
    Issue number19
    DOIs
    StatePublished - Sep 17 1996

    Keywords

    • 5-fluoroorotic acid
    • URA3
    • negative selection

    ASJC Scopus subject areas

    • General

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