Yeast Two-Hybrid System

This volume, part of the Advances in Molecular Biology series, presents detailed protocols and trouble-shooting advice on the yeast two-hybrid system, one of the most powerful and versatile methods for characterizing a protein′s functions. Topics include: characterizing hormone/receptor complexes, identifying peptide ligands, how to identify mutations that disrupt an interaction, how to construct an activation domain hybrid library, and how to dissect the cell cycle and other complex genetic networks.

1. The two-hybrid system: A personal view, S. Fields and P.L. Bartel
I. The Two-Hybrid System
2. Searching for interacting proteins with the two-hybrid system I, C. Bai and S.J. Elledge
3. Searching for interacting proteins with the two-hybrid system II, A.B. Vojtek, J.A. Cooper, and S.M. Hollenberg
4. Searching for interacting proteins with the two-hybrid system III, E.A. Golemis and R. Brent
5. Constructing an activation domain fusion library, L. Zhu, D. Gunn, and S. Kuchibhatla
II. Analyzing Interactions with the Two-Hybrid System
6. Obtaining structural information about protein complexes with the two-hybrid system, D.C. Amberg and D. Botstein
7. The reverse two-hybrid system, M. Vidal
8. Dissecting the biological relevance of multiple interactions, M.A. White
9. Investigation of ligand/receptor interactions and the formation of ternary complexes, B.A. Ozenberger and K.H. Young
10. Two-hybrid analysis of protein-protein interactions in the yeast pheromone response pathway, G.F. Sprague, Jr. and J.A. Printen
11. Two-hybrid screening and the cell cycle, G.J. Hannon
12. Two-hybrid analysis of genetic regulatory networks, R.L. Finley, Jr. and R. Brent
III. Variations on the Two-Hybrid Theme
13. Mammalian cells as hosts for two-hybrid studies of protein-protein interaction, J.T. Tsan, Z. Wang, J. Jin, L.-Y. Hwang, R.O. Bash, and R. Baer
14. Detection of protein-protein interactions dependent on post-translational modifications, M.A. Osborne, M. Lubinus, and J.P. Kochan
15. Protein interactions mediated by small molecule ligands, V. Berlin
16. Use of a combinatorial peptide library in the two-hybrid assay, M. Yang
IV. Related Methodologies
17. Isolation of DNA-binding proteins using a one-hybrid genetic screen, J.A. Chong and G. Mandel
18. A three-hybrid system to detect and analyze RNA-protein interactions in vivo, B. Zhang, B. Kraemer, D. SenGupta, S. Fields, and M. Wickens
19. Split ubiquitin: a sensor of protein interactions in vivo, N. Johnsson and A. Varshavsky