Genetic approaches to the study of protein-protein interactions.
This article describes genetic approaches to the study of heterologous protein-protein interactions, focusing on the yeast Saccharomyces cerevisiae as a useful eukaryotic model system. Several methods are described that can be used to search for new interactions, including extragenic suppression, multicopy suppression, synthetic lethality, and transdominant inhibition. Strategies for screening, genetic characterization, and clone identification are described, along with recent examples from the literature. In addition, genetic methods are discussed that can be used to further characterize a newly discovered protein-protein interaction. These include the creation of mutant libraries of a given protein by chemical mutagenesis or polymerase chain reaction, the production of dominant-negative mutants, and strategies for introducing these mutant alleles back into yeast for analysis. Although these genetic methods are quite powerful, they are often just a starting point for further biochemical or cell biological experiments.
Appling
D R
DR
Department of Chemistry and Biochemistry, The Biochemical Institute, Austin, Texas 78712, USA. dappling@mail.utexas.edu
D003001:Cloning, Molecular / Q000379:methods; D005804:Genes, Lethal; D016153:Genes, Suppressor; D005813:Genes, Synthetic; D005821:Genetic Techniques; D016296:Mutagenesis; D019151:Peptide Library; D017354:Point Mutation; D011485:Protein Binding; D011506:Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism
D019151:Peptide Library; D011506:Proteins; D011994:Recombinant Proteins