Genetic and Biochemical Analysis of the Activation Mechanism of the Saccharomyces Cerevisiae Pheromone Receptor
Authors
Bukusoglu, Gul H.Faculty Advisor
Dr. Duane D. JennessAcademic Program
Molecular Genetics and MicrobiologyUMass Chan Affiliations
Molecular Genetics and MicrobiologyDocument Type
Doctoral DissertationPublication Date
1998-01-28Keywords
Saccharomyces cerevisiaeChemoreceptors
Amino Acids, Peptides, and Proteins
Biological Factors
Fungi
Genetic Phenomena
Metadata
Show full item recordAbstract
Activation mechanism of the α-factor pheromone receptor of Saccharomyces cerevisiae was analyzed using biochemical and genetic techniques. An in vitro partial proteolysis assay was developed to determine the conformational change of the receptor that occurs upon binding of agonist. The activation specific cleavages were established by comparing cleavage products with antagonist versus agonist occupied receptor. Of the changes in peptide pattern that were revealed by trypsinization, the fragment resulting from the exposure of the third loop to the protease was found to be agonist specific and to be G-protein independent. A low-affinity binding receptor mutant was isolated which failed to undergo this agonist induced conformational change. Four intra-allelic suppressors of this receptor mutant were isolated and all were mapped to the ends of transmembrane helices 4, 5, 6 and 7; all were found to be replacements of non-polar residues by polar ones. The role of the suppressor mutations in conformational change was analyzed.DOI
10.13028/1s9w-6e30Permanent Link to this Item
http://hdl.handle.net/20.500.14038/31450Notes
Some images did not scan well. Please consult original document.
Rights
Copyright is held by the author, with all rights reserved.ae974a485f413a2113503eed53cd6c53
10.13028/1s9w-6e30