title: Dissecting a known RNA-protein interaction using a yeast three-hybrid system.
abstract: The yeast three-hybrid system has been applied to known protein-RNA interactions for a variety of purposes. For instance, protein and RNA mutants with altered or relaxed binding specificities can be identified. Mutant RNAs can also be analyzed to better understand RNA-binding specificity of a specific protein. Furthermore, this system complements other biochemical techniques, for example, SELEX, co-immunoprecipitation and cross-linking experiments (see UV crosslinking of interacting RNA and protein in cultured cells and PAR-CLIP (Photoactivatable Ribonucleoside-Enhanced Crosslinking and Immunoprecipitation): a step-by-step protocol to the transcriptome-wide identification of binding sites of RNA-binding proteins).
authors.affiliation: Microbiology Doctoral Training Program, University of Wisconsin - Madison, Madison, WI, USA.
authors.forename: Yvonne Y
authors.initials: YY
authors.lastname: Koh
authors.affiliation: Department of Biochemistry, University of Wisconsin - Madison, Madison, WI, USA. Electronic address: wickens@biochem.wisc.edu.
authors.forename: Marvin
authors.initials: M
authors.lastname: Wickens
mesh_terms: D000640:Amitrole / Q000494:pharmacology; D000935:Antifungal Agents / Q000494:pharmacology; D001665:Binding Sites; D025141:Drug Resistance, Fungal; D017930:Genes, Reporter; D011485:Protein Binding; D012313:RNA / Q000378:metabolism*; D016601:RNA-Binding Proteins / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000187:drug effects / Q000254:growth & development; D014170:Transformation, Genetic; D020798:Two-Hybrid System Techniques; D001616:beta-Galactosidase / Q000096:biosynthesis / Q000235:genetics
chemical_list: D000935:Antifungal Agents; D016601:RNA-Binding Proteins; D012313:RNA; D001616:beta-Galactosidase; D000640:Amitrole
keywords: 3–Aminotriazole (3–AT) resistance assay; DNA-binding site; Plasmids; RNA–protein interaction; Yeast three-hybrid system; β-Galactosidase activity