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Quantitative analysis of the yeast pheromone pathway
Author(s) -
Shellhammer James P.,
Pomeroy Amy E.,
Li Yang,
Dujmusic Lorena,
Elston Timothy C.,
Hao Nan,
Dohlman Henrik G.
Publication year - 2019
Publication title -
yeast
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.923
H-Index - 102
eISSN - 1097-0061
pISSN - 0749-503X
DOI - 10.1002/yea.3395
Subject(s) - biology , saccharomyces cerevisiae , yeast , computational biology , model organism , pheromone , population , mapk/erk pathway , phosphorylation , microbiology and biotechnology , genetics , gene , demography , sociology
The pheromone response pathway of the yeast Saccharomyces cerevisiae is a well‐established model for the study of G proteins and mitogen‐activated protein kinase (MAPK) cascades. Our longstanding ability to combine sophisticated genetic approaches with established functional assays has provided a thorough understanding of signalling mechanisms and regulation. In this report, we compare new and established methods used to quantify pheromone‐dependent MAPK phosphorylation, transcriptional induction, mating morphogenesis, and gradient tracking. These include both single‐cell and population‐based assays of activity. We describe several technical advances, provide example data for benchmark mutants, highlight important differences between newer and established methodologies, and compare the advantages and disadvantages of each as applied to the yeast model. Quantitative measurements of pathway activity have been used to develop mathematical models and reveal new regulatory mechanisms in yeast. It is our expectation that experimental and computational approaches developed in yeast may eventually be adapted to human systems biology and pharmacology.