| Title: | Reagents for investigating MAPK signalling in model yeast species |
| Author(s): | Pincus D; Benjamin K; Burbulis I; Tsong AE; Resnekov O; |
| Address: | "Molecular Sciences Institute, 2168 Shattuck Avenue, Berkeley, CA 94704, USA" |
| ISSN/ISBN: | 1097-0061 (Electronic) 0749-503X (Linking) |
| Abstract: | "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" |
| Keywords: | "Animals Antibodies, Fungal/*immunology/isolation & purification Fungal Proteins/genetics/isolation & purification/*metabolism Gene Expression *Gene Expression Regulation, Fungal Genetic Vectors Kluyveromyces/metabolism/physiology Mitogen-Activated Protein;" |
| Notes: | "MedlinePincus, David Benjamin, Kirsten Burbulis, Ian Tsong, Annie E Resnekov, Orna eng P50 HG002370/HG/NHGRI NIH HHS/ Research Support, N.I.H., Extramural England 2010/02/18 Yeast. 2010 Jul; 27(7):423-30. doi: 10.1002/yea.1758" |
