title: Using the yeast interaction trap and other two-hybrid-based approaches to study protein-protein interactions.
abstract: The detection of physical interaction between two or more molecules of interest can be facilitated if the act of association between the interactive partners leads to the production of a readily observed biological or physical readout. Many interacting molecule pairs (X, Y) can be made to induce such a readout if X and Y are each fused to defined protein elements with desired properties. For example, in the yeast forward two-hybrid system, X is synthesized as a translational fusion to a DNA-binding domain (DBD), Y is synthesized as a fusion to a transcriptional activation domain (AD), and coexpression of DBD-X and AD-Y induces transcription of easily scored responsive reporters. Other approaches use paradigms based on the artificial production of two, hybrid, molecules, but substitute a variety of readouts including the repression of transcription, activation of signal transduction pathways, or reconstitution of a disrupted enzymatic activity. In this article, we summarize a number of two-hybrid-based approaches, and detail the use of the forward yeast two-hybrid system in a screen to identify novel interacting partners for a protein of interest.
authors.affiliation: Division of Basic Science, Fox Chase Cancer Center, Philadelphia, Pennsylvania 19111, USA.
authors.forename: G G
authors.initials: GG
authors.lastname: Toby
authors.forename: E A
authors.initials: EA
authors.lastname: Golemis
mesh_terms: D004274:DNA, Recombinant / Q000737:chemistry / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000378:metabolism; D015723:Gene Library; D005822:Genetic Vectors / Q000378:metabolism; D066298:In Vitro Techniques; D019151:Peptide Library; D010957:Plasmids / Q000378:metabolism; D011506:Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D012097:Repressor Proteins / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics; D014158:Transcription, Genetic / Q000502:physiology; D014170:Transformation, Genetic / Q000502:physiology; D020798:Two-Hybrid System Techniques
chemical_list: D004274:DNA, Recombinant; D004268:DNA-Binding Proteins; D019151:Peptide Library; D011506:Proteins; D012097:Repressor Proteins