Yeast Two‐hybrid System and Related Methodology

Russell L Jr Finley, Wayne State University School of Medicine, Detroit, Michigan, USA

Published online: January 2006

DOI: 10.1038/npg.els.0005980

The identification and detailed characterization of the binary interactions between gene products in a cell is an essential step toward understanding the functions of genes and how they control cellular systems. Two-hybrid assays have become powerful tools for this purpose.

Keywords: two-hybrid; yeast; protein interaction; networks; complementation

Figure 1. Two-hybrid assays for protein–protein interactions. In a two-hybrid assay, each protein to be tested is expressed fused to a tag, A or B. In this example, X interacts with Y (right) but not Y¢ (left). When A and B are brought near each other through the X–Y interaction, they activate a signal that can be detected (right). The signal may result from direct interaction between A and B, for example, if together A and B form an active enzyme. Alternatively, the signal may be generated when one tag becomes localized to a particular subcellular location as a result of the interaction, as in transcription-based two-hybrid systems.
Figure 2. In the transcription-based yeast two-hybrid system, one tag is a DNA-binding domain (DBD) which binds to specific sites in a reporter gene. The other tag is a transcription-activation domain (AD). An interaction between X and Y localizes the AD to the reporter, where it activates transcription.
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 References
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 Further Reading
    book Bartel PL and Fields S (1997) "The two-hybrid system: a personal view". In: Bartel PL and Fields S (eds.) The Yeast Two-hybrid System, pp. 3–7. Oxford, UK: Oxford University Press.
    Brent R and Finley RL Jr (1997) Understanding gene and allele function with two-hybrid methods. Annual Review of Genetics 31: 663–704.
    book Golemis EA (2002) Protein–Protein Interactions. Plainview, NY: Cold Spring Harbor Laboratory Press.
    Hazbun TR and Fields S (2001) Networking proteins in yeast. Proceedings of the National Academy of Sciences of the United States of America 98: 4277–4278.
    Hu JC, Kornacker MG and Hochschild A (2000) Escherichia coli one- and two-hybrid systems for the analysis and identification of protein–protein interactions. Methods (San Diego, CA) 20: 80–94.
    Ito T, Chiba T, Ozawa R, et al. (2001) A comprehensive two-hybrid analysis to explore the yeast protein interactome. Proceedings of the National Academy of Sciences of the United States of America 98: 4569–4574.
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    Rain JC, Selig L, De Reuse H, et al. (2001) The protein–protein interaction map of Helicobacter pylori. Nature 409: 211–215.
    Remy I and Michnick SW (2001) Visualization of biochemical networks in living cells. Proceedings of the National Academy of Sciences of the United States of America 98: 7678–7683.
    Uetz P, Giot L, Cagney G, et al. (2000) A comprehensive analysis of protein–protein interactions in Saccharomyces cerevisiae. Nature 403: 623–627.
    Walhout AJ and Vidal M (2001) Protein interaction maps for model organisms. Nature Reviews Molecular Cell Biology 2: 55–62.

Related Sub-Topics:

Techniques in Cell Biology | Molecular Genetics of Common and Complex Disease | Techniques and Tools in Clinical Genetics | Posttranslational Modification | Techniques and Tools in Molecular Biology
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Article Title: Yeast Two‐hybrid System and Related Methodology
 
How to Cite close
Finley, Russell L Jr(Jan 2006) Yeast Two‐hybrid System and Related Methodology. In: eLS. John Wiley & Sons Ltd, Chichester. http://www.els.net [doi: 10.1038/npg.els.0005980]