A novel genetic system to detect protein–protein interactions (original) (raw)

Nature volume 340, pages 245–246 (1989)Cite this article

Abstract

PROTEIN-protein interactions between two proteins have generally been studied using biochemical techniques such as crosslinking, co-immunoprecipitation and co-fractionation by chromatography. We have generated a novel genetic system to study these interactions by taking advantage of the properties of the GAL4 protein of the yeast Saccharomyces cerevisiae. This protein is a transcriptional activator required for the expression of genes encoding enzymes of galactose utilization1. It consists of two separable and functionally essential domains: an N-terminal domain which binds to specific DNA sequences (UASG); and a C-terminal domain containing acidic regions, which is necessary to activate transcription2,3. We have generated a system of two hybrid proteins containing parts of GAL4: the GAL4 DNA-binding domain fused to a protein 'X' and a GAL4 activating region fused to a protein 'Y'. If X and Y can form a protein-protein complex and reconstitute proximity of the GAL4 domains, tran-scription of a gene regulated by UASG occurs. We have tested this system using two yeast proteins that are known to interact—SNF1 and SNF4. High transcriptional activity is obtained only when both hybrids are present in a cell. This system may be applicable as a general method to identify proteins that interact with a known protein by the use of a simple galactose selection.

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Authors and Affiliations

  1. Department of Microbiology, State University of New York at Stony Brook, Stony Brook, New York, 11794, USA
    Stanley Fields & Ok-kyu Song

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  1. Stanley Fields
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  2. Ok-kyu Song
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Fields, S., Song, Ok. A novel genetic system to detect protein–protein interactions.Nature 340, 245–246 (1989). https://doi.org/10.1038/340245a0

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