Functional complementation of yeast mutants to study plant signalling pathways.
The rapidly increasing amount of entirely sequenced genomes generates a need for fast and efficient methods to elucidate gene functions. Functional complementation of yeast mutants, displaying selectable phenotypes, has been used very successfully in the past years to isolate many plant genes involved in signalling, stress response or metabolic pathways. Using the well-characterized Hog1 pathway, a mitogen activated protein (MAP) kinase pathway required for adaptation to osmotic stress in budding yeast, as example, we describe here the isolation of plant protein kinases involved in abiotic stress adaptation in the model plant Arabidopsis thaliana. The osmo-sensitive phenotype of yeast mutants carrying a mutation in the Hog1 pathway allows an easy selection on high osmolarity media, containing i.e. 0.4 M NaCl. By using yeast mutants harbouring deletions in different components of the pathway, for example the MAP kinase kinase Pbs2 and the MAP kinase Hog1, it is furthermore possible to isolate consecutively acting components of a signalling pathway.
Mehlmer
Norbert
N
Department of Biochemistry, Max F. Perutz Laboratories, University of Vienna, Vienna, Austria.
Scheikl-Pourkhalil
Elisabeth
E
Teige
Markus
M
D017360:Arabidopsis / Q000235:genetics / Q000378:metabolism; D005816:Genetic Complementation Test; D020930:MAP Kinase Kinase Kinases / Q000235:genetics / Q000378:metabolism; D020928:Mitogen-Activated Protein Kinases / Q000235:genetics / Q000378:metabolism; D008957:Models, Genetic; D009154:Mutation; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism / Q000502:physiology; D010944:Plants / Q000235:genetics / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction / Q000235:genetics / Q000502:physiology*; D015003:Yeasts / Q000235:genetics / Q000378:metabolism*
D010940:Plant Proteins; D029701:Saccharomyces cerevisiae Proteins; C088589:HOG1 protein, S cerevisiae; D020928:Mitogen-Activated Protein Kinases; D020930:MAP Kinase Kinase Kinases