Premium
Reagents for investigating MAPK signalling in model yeast species
Author(s) -
Pincus David,
Benjamin Kirsten,
Burbulis Ian,
Tsong Annie E.,
Resnekov Orna
Publication year - 2010
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.1758
Subject(s) - kluyveromyces lactis , biology , saccharomyces cerevisiae , schizosaccharomyces pombe , polyclonal antibodies , mapk/erk pathway , yeast , microbiology and biotechnology , plasmid , antibody , signal transduction , genetics , gene
Here we present a set of resources (bacterial expression plasmids and antibodies) for the interrogation of proteins involved in yeast MAPK signalling. We constructed bacterial protein expression plasmids for 25 proteins involved in MAPK signalling in budding yeast. From these constructs we expressed and purified proteins and generated rabbit polyclonal antibodies against 13 proteins in the pheromone MAPK pathway. We verified the specificity of the antibodies and employed them to follow pathway proteins in cells stimulated with pheromone. We show that these reagents can be used to detect pheromone‐induced post‐translational modifications and changes in the oligomeric state of pathway proteins. In addition to recognizing their target proteins in Saccharomyces cerevisiae , these antibodies allow the detection of predicted orthologues in the distant evolutionary relatives Kluyveromyces lactis and Schizosaccharomyces pombe . These antibodies are new tools for investigating MAPK signalling in model yeast species and may be useful for studying MAPK signalling in higher eukaryotes. Copyright © 2010 John Wiley & Sons, Ltd.