| Title: | Systematic mutagenesis of the yeast mating pheromone receptor third intracellular loop |
| Author(s): | Clark CD; Palzkill T; Botstein D; |
| Address: | "Department of Genetics, Stanford University School of Medicine, California 94305" |
| ISSN/ISBN: | 0021-9258 (Print) 0021-9258 (Linking) |
| Abstract: | "Signal transduction in the mating pathway of the yeast Saccharomyces cerevisiae is initiated by binding of a peptide pheromone to a G protein-coupled receptor (Ste2). We systematically have mutated the third intracellular loop of the Ste2 receptor to investigate its functional significance. We substituted each of the 13 amino acids in the loop with alanine individually or together with one other residue. In addition, we used a site-directed random replacement mutagenesis technique to replace a region encoding three amino acids in the loop with random sequence. Over 80 such Ste2 mutants have been analyzed by several functional and biochemical criteria in a yeast strain that carries a genomic deletion of the STE2 gene. The mutant phenotypes range from fully functional to severely compromised in signaling. The observation that amino acid substitutions in the third intracellular loop of the Ste2 receptor can affect activation of the yeast mating response implicates the loop in this signal transduction pathway. The types of mutations that compromise the function of the receptor may provide clues to the physical interaction between the receptor and the G protein" |
| Keywords: | "Amino Acid Sequence Cytoplasm/metabolism Fungal Proteins/chemistry GTP-Binding Proteins/physiology Mating Factor Molecular Sequence Data Mutagenesis, Site-Directed Peptides/physiology Receptors, Mating Factor Receptors, Peptide/*chemistry Saccharomyces ce;" |
| Notes: | "MedlineClark, C D Palzkill, T Botstein, D eng GM46406/GM/NIGMS NIH HHS/ GM46888/GM/NIGMS NIH HHS/ Comparative Study Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. 1994/03/25 J Biol Chem. 1994 Mar 25; 269(12):8831-41" |
