Yeast three-hybrid

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. 

Gardiner L, Coyle BJ, Chan WC, Soultanas P. Discovery of antagonist peptides against bacterial helicase-primase interaction 79. in B. stearothermophilus by reverse yeast three-hybrid. Chem. 

Henthorn DC, Jaxa-Chamiec AA, Meldmm E. A GAL4-based yeast three-hybrid system for the identification of small molecule-target protein interactions. Biochem. Pharmacol. 2002 63 1619-1628. 

SenGupta DJ, Zhang B, Kraemer B, Pochart P, Fields S, Wickens M. (1996). A three-hybrid system to detect RNA-protein interactions in vivo. Proc. Natl. Acad. Sci. U.S.A. 1996 93 8496-8501. Jaeger S, Eriani G, Martin F. Results and prospects of the yeast three-hybrid system. FEES Lett. 2004 556 7-12. 

Konig J, Julius C, Baumann S, Homann M, Goringer HU, Feldbmgge M. Combining SELEX and the yeast three-hybrid system for in vivo selection and classification of RNA aptamers. RNA 2007 13 614-622. 

Riley KJ, Cassiday LA, Kumar A, Maher LJ 3rd. Recognition of RNA by the p53 tumor suppressor protein in the yeast three-hybrid system. Rna 2006 12 620-630. 

Bernstein DS, Buter N, Smmpf C, Wickens M. Analyzing mRNA-protein complexes using a yeast three-hybrid system. Methods 2002 26 123-141. 

Martin F, Michel F, Zenklusen D, Muller B, Schumperli D. Positive and negative mutant selection in the human histone hairpin-binding protein using the yeast three-hybrid system. Nucleic Acids Res. 2000 28 1594-1603. 

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. 

Hussey SL, Muddana SS, Peterson BR. Synthesis of a beta-estradiol-biotin chimera that potently heterodimerizes estrogen receptor and streptavidin proteins in a yeast three-hybrid system. J. Am. Chem. Soc. 2003 125 3692-3693. 

Just as a dimeric RNA molecule can be introduced to mediate the interaction between the DNA-binding and ADs, so can a dimeric small molecule [17]. In fact, well before their use in a small molecule three-hybrid assay, dimeric small molecules were used as chemical inducers of dimerization (CIDs) to artificially oligomerize fusion proteins in vivo [18]. In the yeast three-hybrid system, the union of two protein fusions and a CID reconstitute the transcription of a reporter gene (Fig. 4.1-3(d)). In 1996, Licitra and Liu built what they called a yeast three-hybrid assay [19]. This assay consists of two fusion proteins and a heterodimeric small molecule CID that brings these fusion proteins together to activate the transcription of a reporter gene (Fig. 4.1-3 d)). 

Fig. 4.1-5 Small molecules used to create chemical inducers of dimerization (Cl Ds) for the yeast three-hybrid system.

S. Jaeger, G. Eriani, F. Martin, Results and prospects of the yeast three-hybrid system, FEBS Lett. 2004, 556, 7-12. 

S. Fields, M. Wickens, Yeast three-hybrid system to detect and analyze interactions between RNA and protein, Methods Enzymol. 1999, 306, 93-113. 

S.S. Muddana, B.R. Peterson, Facile synthesis of cids biotinylated estrone oximes efficiently heterodimerize estrogen receptor and streptavidin proteins in yeast three hybrid systems, Org. Lett. 2004, 6, 1409-1412. 

K.S. de Felipe, B.T. Carter, E.A. Althoff, V.W. Cornish, Correlation between ligand-receptor affinity and the transcription readout in a yeast three-hybrid system, Biochemistry 2004, 43,10353-10363. 

The yeast three-hybrid (Y3H) system is a cellular assay system designed for the identification and characterization of small molecule-protein interactions in intact cells [25]. It uses yeast Saccharomyces cerevisiae as a host system and combines aspects of the yeast two-hybrid (Y2H) system [26] with recent developments in chemical dimerizer technology [27, 28],... 

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... 

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