Gg. Toby et Ea. Golemis, Using the yeast interaction trap and other two-hybrid-based approaches to study protein-protein interactions, METHODS, 24(3), 2001, pp. 201-217
The detection of physical interaction between two or more molecules of inte
rest can be facilitated if the act of association between the interactive p
artners leads to the production of a readily observed biological or physica
l readout. Many interacting molecule pairs (X, Y) can be made to induce suc
h a readout if X and Y are each fused to defined protein elements with desi
red properties. For example, in the yeast forward two-hybrid system, X is s
ynthesized as a translational fusion to a DNA-binding domain (DBD), Y is sy
nthesized as a fusion to a transcriptional activation domain (AD), and coex
pression of DBD-X and AD-Y induces transcription of easily scored responsiv
e reporters. Other approaches use paradigms based on the artifical producti
on of two, hybrid, molecules, but substitute a variety of readouts includin
g the repression of transcription, activation of signal transduction pathwa
ys, or reconstitution of a disrupted enzymatic activity. In this article, w
e 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 interacti
ng partners for a protein of interest. (C) 2001 Academic Press.