Analyzing mRNA-protein complexes using a yeast three-hybrid system.

David S Bernstein; Natascha Buter; Craig Stumpf; Marvin Wickens

doi:10.1016/s1046-2023(02)00015-4

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Analyzing mRNA-protein complexes using a yeast three-hybrid system.

David S Bernstein·Natascha Buter·Craig Stumpf·Marvin Wickens

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Abstract

RNA-protein interactions are essential for the proper execution and regulation of every step in the life of a eukaryotic mRNA. Here we describe a three-hybrid system in which RNA-protein interactions can be analyzed using simple phenotypic or enzymatic assays in Saccharomyces cerevisiae. The system can be used to detect or confirm an RNA-protein interaction, to analyze RNA-protein interactions genetically, and to discover new protein or RNA partners when only one is known. Multicomponent complexes containing more than one protein can be detected, identified, and analyzed. We describe the method and how to use it, and discuss applications that bear particularly on eukaryotic mRNAs.

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Amitrole Binding Sites DNA, Complementary Enzyme Inhibitors Gene Library Ligands Models, Genetic Nucleic Acid Conformation Phenotype Protein Binding Protein Structure, Tertiary RNA RNA, Messenger RNA-Binding Proteins Saccharomyces cerevisiae Two-Hybrid System Techniques beta-Galactosidase Journal Article Research Support, U.S. Gov't, P.H.S.

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David S Bernstein, Natascha Buter, Craig Stumpf, Marvin Wickens. 2002. Analyzing mRNA-protein complexes using a yeast three-hybrid system.. https://doi.org/10.1016/s1046-2023(02)00015-4

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