Two-hybrid and Related Systems

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Two-hybrid and Related Systems

Stanley Fields, University of Washington, Seattle, Washington, USA

Published online: April 2001

DOI: 10.1038/npg.els.0000981

Full Article on Wiley Online Library

Abstract
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References

The two-hybrid system is a yeast-based genetic method to detect and analyse protein–protein interactions. Related systems have been developed to detect DNA–protein, RNA–protein and small molecule–protein interactions.

Keywords: protein–protein interaction; DNA–protein interaction; RNA–protein interaction; yeast; transcription factor

References
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Further Reading
	book Bartel PL and Fields S (eds) (1997) The Yeast Two-Hybrid System. Oxford: Oxford University Press.
	Brachman RK and Boeke JD (1997) Tag games in yeast: the two-hybrid system and beyond. Current Opinion in Biotechnology 8: 561–568.
	Brent R and Finley RL Jr (1997) Understanding gene and allele function with two-hybrid methods. Annual Review of Genetics 31: 663–704.
	Frederickson RM (1998) Macromolecular matchmaking: advances in two-hybrid and related technologies. Current Opinion in Biotechnology 9: 90–96.
	McNabb DS and Guarente L (1996) Genetic and biochemical probes for protein–protein interactions. Current Opinion in Biotechnology 7: 554–559.

Article Title:	Two-hybrid and Related Systems
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	Figure 1. The two-hybrid system. (a) A transcriptional activator may contain a DNA-binding domain (filled circle) and a transcription activation domain (filled rectangle). (b) A hybrid protein composed of a DNA-binding domain and a protein ‘X’ does not activate transcription if X does not have an activation domain. (c) A hybrid protein composed of an activation domain and a protein ‘Y’ does not activate transcription because it does not bind to the DNA-binding sites. (d) Interaction between X and Y reconstitutes the function of the activator and leads to expression of the reporter gene.
	Figure 2. The three-hybrid system to detect RNA–protein interactions. A hybrid protein of the LexA protein fused to the bacteriophage MS2 coat protein localizes to the regulatory region of the reporter gene by binding to the LexA-binding sites. A hybrid protein of the Gal4 transcription activation domain fused to a second RNA-binding protein will activate transcription of the reporter gene when in close proximity to the gene's regulatory region. A hybrid RNA containing cognate binding sites for the coat protein and the other RNA-binding protein bridges the two hybrid proteins and results in expression of the reporter gene.
	Figure 3. The three-hybrid system to detect the interaction of a protein with a small molecule. A hybrid protein of a DNA-binding domain with a protein domain that binds to ligand A, and a hybrid protein of an activation domain with a protein domain that binds to ligand B, are bridged by a heterodimer of A linked to B.

Two-hybrid and Related Systems

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