title: Identification of kinase substrates by bimolecular complementation assays.
abstract: As a consequence of the transient nature of kinase-substrate interactions, the detection of kinase targets, although central for understanding many biological processes, has remained challenging. Here we present a straightforward procedure that relies on the comparison of wild type with activation-loop mutants in the kinase of interest by bimolecular complementation assays. As a proof of functionality, we present the identification and in vivo confirmation of substrates of the major cell-cycle kinase in Arabidopsis, revealing a direct link between cell proliferation and the control of the redox state.
authors.affiliation: Unigruppe am Max-Planck-Institut für Züchtungsforschung, Max-Delbrück-Laboratorium, Lehrstuhl für Botanik III, Universität Köln, Carl-von-Linné-Weg 10, D-50829 Köln, Germany.
authors.forename: Stefan
authors.initials: S
authors.lastname: Pusch
authors.forename: Hirofumi
authors.initials: H
authors.lastname: Harashima
authors.forename: Arp
authors.initials: A
authors.lastname: Schnittger
mesh_terms: D017360:Arabidopsis / Q000166:cytology / Q000235:genetics / Q000378:metabolism*; D029681:Arabidopsis Proteins / Q000235:genetics / Q000378:metabolism*; D049109:Cell Proliferation; D018844:Cyclin-Dependent Kinases / Q000235:genetics / Q000378:metabolism*; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D018613:Microscopy, Confocal; D009154:Mutation; D010084:Oxidation-Reduction; D011485:Protein Binding; D025941:Protein Interaction Mapping / Q000379:methods*; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000254:growth & development / Q000378:metabolism; D013379:Substrate Specificity; D020798:Two-Hybrid System Techniques
chemical_list: D029681:Arabidopsis Proteins; D008164:Luminescent Proteins; C434472:CDKA1 protein, Arabidopsis; D018844:Cyclin-Dependent Kinases