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07:59
Temporal Quantification of MAPK Induced Expression in Single Yeast Cells
Published on: October 4, 2013
Reagents for investigating MAPK signalling in model yeast species
David Pincus1, Kirsten Benjamin, Ian Burbulis
1Molecular Sciences Institute, 2168 Shattuck Avenue, Berkeley, CA 94704, USA.
Yeast (Chichester, England)
|February 18, 2010
Summary
Researchers developed bacterial expression plasmids and antibodies to study yeast mitogen-activated protein kinase (MAPK) signaling pathways. These new tools enable detection of pathway protein modifications and interactions in various yeast species.
Area of Science:
- Molecular Biology
- Cell Signaling
- Yeast Genetics
Background:
- Mitogen-activated protein kinase (MAPK) signaling pathways regulate crucial cellular processes in yeast.
- Investigating yeast MAPK pathways requires specific tools to detect protein dynamics and modifications.
Purpose of the Study:
- To generate and validate novel reagents, including bacterial expression plasmids and antibodies, for studying yeast MAPK signaling.
- To enable the detection of pheromone-induced changes in MAPK pathway proteins in Saccharomyces cerevisiae.
Main Methods:
- Construction of bacterial expression plasmids for 25 yeast MAPK proteins.
- Expression, purification, and antibody generation against 13 pheromone MAPK pathway proteins.
- Antibody specificity verification and application in pheromone-stimulated yeast cells.
Main Results:
- Successfully generated and validated polyclonal antibodies against 13 MAPK pathway proteins.
- Demonstrated antibody utility in detecting pheromone-induced post-translational modifications and oligomeric state changes.
- Showcased antibody cross-reactivity with orthologs in Kluyveromyces lactis and Schizosaccharomyces pombe.
Conclusions:
- Developed a valuable set of resources (plasmids and antibodies) for yeast MAPK signaling research.
- These antibodies serve as effective tools for investigating MAPK signaling in model yeast species.
- The reagents may also find applications in studying MAPK signaling in higher eukaryotes.
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