International Mammalian Genome Society

logo18th International Mouse Genome Conference

17-22 October 2004, Seattle, USA


POSTER 106- HIGH THROUGHPUT YEAST TWO-HYBRID STRATEGIES FOR PROTEOME-WIDE PROTEIN-PROTEIN INTERACTION MAPS

Zhong J 1, Finley R 2

1 Current affiliation: Laboratory of Genetics, National Institute on Aging, NIH,, Baltimore, MD, United States,

2 Center for Molecular Medicine and Genetics, Wayne State University, Detroit, MI, United States

Most cellular activities are carried out by stable or transient protein-protein interactions. A proteome-wide protein interaction map could suggest the functions of individual proteins, help map entire pathways or processes, and suggest how individual pathways and processes are integrated into larger cellular events. The yeast two-hybrid system is an efficient and sensitive way to detect binary protein interactions, and can be modified for high throughput assays. However, most high throughput two-hybrid approaches fail to detect many interactions that should be detected by yeast two-hybrid systems and they produce a significant number of false positives. We developed an alternative high throughput two-hybrid approach  in which two yeast arrays, conditionally expressing AD or BD fusions, are mated using a two-phase pooled mating scheme. We show that our approach allows us to detect interactions not detected by other approaches, including interactions involving proteins which can activate the reporters on their own, and proteins that are toxic to yeast. We demonstrated that our approach is efficient and can be applied to complex organisms such as Drosophila. Combined, our results suggest that multiple studies using different approaches and different systems may be necessary to uncover all interactions that can be detected by yeast two hybrid assays.

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