Schlecht U, et al. (2012) Multiplex assay for condition-dependent changes in protein-protein interactions. Proc Natl Acad Sci U S A 109(23):9213-8
Abstract: Changes in protein-protein interactions that occur in response to environmental cues are difficult to uncover and have been poorly characterized to date. Here we describe a yeast-based assay that allows many binary protein interactions to be assessed in parallel and under various conditions. This method combines molecular bar-coding and tag array technology with the murine dihydrofolate reductase-based protein-fragment complementation assay. A total of 238 protein-fragment complementation assay strains, each representing a unique binary protein complex, were tagged with molecular barcodes, pooled, and then interrogated against a panel of 80 diverse small molecules. Our method successfully identified specific disruption of the Hom3:Fpr1 interaction by the immunosuppressant FK506, illustrating the assay's capacity to identify chemical inhibitors of protein-protein interactions. Among the additional findings was specific cellular depletion of the Dst1:Rbp9 complex by the anthracycline drug doxorubicin, but not by the related drug idarubicin. The assay also revealed chemical-induced accumulation of several binary multidrug transporter complexes that largely paralleled increases in transcript levels. Further assessment of two such interactions (Tpo1:Pdr5 and Snq2:Pdr5) in the presence of 1,246 unique chemical compounds revealed a positive correlation between drug lipophilicity and the drug response in yeast.
|Status: Published||Type: Journal Article||PubMed ID: 22615397|
Topics addressed in this paper
Number of different genes curated to this paper: 11
- To go to the Locus page for a gene, click on the gene name.
|Topics||Topics not linked to Genes||Genes (#1 - 10 )|
|Large-scale protein interaction|
|Techniques and Reagents|
|Topics||Genes (#11 )|