Identification of constitutively active AtMPK6 mutants using a functional screen in Saccharomyces cerevisiae

Elodie Hudik; Souha Berriri; Heribert Hirt; Jean Colcombet

doi:10.1007/978-1-4939-0922-3_6

Identification of constitutively active AtMPK6 mutants using a functional screen in Saccharomyces cerevisiae

Methods Mol Biol. 2014:1171:67-77. doi: 10.1007/978-1-4939-0922-3_6.

Authors

Elodie Hudik¹, Souha Berriri, Heribert Hirt, Jean Colcombet

Affiliation

¹ Institut de Biologie des Plantes, UMR CNRS-8618, Centre National De La Recherche Scientifique, Rue de Noetzlin, Bâtiment 630, 91405, Orsay Cedex, France.

PMID: 24908120
DOI: 10.1007/978-1-4939-0922-3_6

Abstract

MAPK (Mitogen-Activated Protein Kinases) mutants which are active independently of phosphorylation by upstream MAPK Kinases (MAPKKs) help to clarify signal transduction processes through MAPK modules and provide a useful tool to understand MAPK roles in the cell. The identification of such mutations is tricky. In this chapter, we provide a detailed protocol for their screening, taking advantage of a functional expression assay in yeast.

MeSH terms

Arabidopsis / enzymology*
Arabidopsis / genetics
Arabidopsis Proteins / genetics*
Genetic Engineering / methods*
Mitogen-Activated Protein Kinases / genetics*
Mutation*
Plasmids / genetics
Saccharomyces cerevisiae / drug effects
Saccharomyces cerevisiae / genetics*
Saccharomyces cerevisiae / growth & development
Salts / pharmacology
Transformation, Genetic

Substances

Arabidopsis Proteins
Salts
MPK6 protein, Arabidopsis
Mitogen-Activated Protein Kinases