Three hybrid screens

Combining Two- and Three-Hybrid Screens with Other Technoiogies... 

The hrst step toward this goal is to document the interactions. To this end, powerful tools have been developed to scan for macromolecular interactions. Arguably, the most significant and far-reaching of these methods is the yeast two-hybrid screen (1). Subsequent work has produced a host of thematic variations including the three-hybrid screen, a method first described explicitly in 1996 that employs small molecules or nucleic acids as baits (2). In this review, we discuss some strengths and... 

Figure 4 Architecture of three-hybrid screens, (a) The classic, small-molecule-based, three-hybrid platform is shown. The three components of this systems are a DNA-binding protein fused to a known drug-binding domain (X), a bifunctional small molecule that binds X on one end and displays a query "bait" on the other, and a library of possible "prey" proteins (V) expressed as fusions to transcriptional activation domains. Assembly of the ternary complex recruits the activation domain to the promoter and initiates reporter gene expression. A partial list of examples is shown in the inset boxes because of space constraints, some relevant systems are absent, (b) The RNA-based, three-hybrid is assembled in a similar fashion except that the bifunctional ligand is an RNA molecule. The complex between the MS2 RNA and the MS2-coat protein is typically used as the anchoring interaction.

S. L. Schreiber, Inducible gene expression and protein translocation using nontoxic ligands identified by a mammalian three-hybrid screen, Proc. 

S.D. Liberies, S.T. Diver, D.J. Austin, S.L. Schreiber, Inducible gene expression and protein translocation using nontoxic ligands identified by a mammalian three-hybrid screen, Proc. Nall. Acad. Sci. U.S.A. 1997, 94, 7825-7830. 

There are two other technologies stiU in development with similar potentials. So-called yeast three hybrid screens are related to the better known two-hybrid screens, where the binding interaction that is being hunted for triggers the assembly of a dimeric transcription factor. The functional transcription factor then stimulates expression of a reporter gene. One half of the transcription factor is fused to a known smaU-molecule receptor (say for a dmg), whilst the other is fused to memhers... 

Shepard, A.R., Conrow, R.E., Pang, I.-H. et al. (2013) Identification of PDE6D as a molecular target of anecortave acetate via a methotrexate-anchored yeast three-hybrid screen. ACS Chemical Biology, 8, 549-558. 

PIAS (protein inhibitors of activated STATs) proteins were first discovered in yeast-two-hybrid screens as interacting molecules with STAT transcription factors. The mammalian family consists ofthe founding member PIAS3, which was described as a repressor of STAT3, and three additional members, PIAS1, PIASy (also known as PIAS4), and PIASx (also known as... 

In general, protein target identification often employs genetic techniques such as expression cloning, expression profiling, screening of yeast mutations, and yeast three-hybrid assays. None of these techniques works for every situation. 

A wide variety of library screening technologies have been developed. Two and three hybrid systems and analogous technologies have proved extremely useful for detecting pairs of interacting proteins or peptides... 

Hook B, Bernstein D, Zhang B, Wickens M. RNA-protein interactions in the yeast three-hybrid system affinity, sensitivity, and enhanced library screening. Rna 2005 11 227-233. 

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