Tag games in yeast: the two-hybrid system and beyond

• Published in: Current opinion in biotechnology Vol. 8; no. 5; pp. 561 - 568
• Main Authors: Brachmann, Rainer K, Boeke, Jef D
• Format: Journal Article
• Language: English
• Published: England Elsevier Ltd 01.10.1997
• Subjects: Animals; Biotechnology - methods; Cloning, Molecular - methods; Genes, Reporter; Humans; Models, Genetic; Recombinant Fusion Proteins - biosynthesis; Recombinant Proteins - biosynthesis; Saccharomyces cerevisiae - genetics; Saccharomyces cerevisiae - physiology; CREB cAMP-response element binding protein; DBD DNA binding domain; GEF guanyl nucleotide exchange factor 1HS one-hybrid system; 3HS three-hybrid system; UAS upstream activating sequence; CID chemical inducer of dimerization; TetR tet-repressor; AD activation domain; 2HS two-hybrid system; 5-Foa 5-fluoroorotic acid; R2HS reverse 2HS; RRE Rev responsive element; SRS Sos recruitment system; USPS ubiquitin-based split-protein sensor; UBP ubiquitin-specific protease
• ISSN: 0958-1669; 1879-0429
• DOI: 10.1016/S0958-1669(97)80029-8

The yeast Saccharomyces cerevisiae and the one- and two-hybrid systems are essential genetic tools of studying the macromolecular interactions that define all living organisms. Newly developed variations on this theme can now address an even bigger set of questions. Reverse one- and two-hybrid systems can identify factors that dissociate or abrogate defined macromolecular interactions. Different forms of three-hybrid systems can evaluate the complex interplay of proteins with RNAs, peptide ligands, small organic ligands or protein kinases. Finally, the ubiquitin-based split-protein sensor and the Sos recruitment systems promise to overcome some limitations of conventional two-hybrid systems.