Identification of kinase substrates by bimolecular complementation assays

Plant J. 2012 Apr;70(2):348-56. doi: 10.1111/j.1365-313X.2011.04862.x. Epub 2012 Jan 10.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Arabidopsis / cytology
  • Arabidopsis / genetics
  • Arabidopsis / metabolism*
  • Arabidopsis Proteins / genetics
  • Arabidopsis Proteins / metabolism*
  • Cell Proliferation
  • Cyclin-Dependent Kinases / genetics
  • Cyclin-Dependent Kinases / metabolism*
  • Luminescent Proteins / genetics
  • Luminescent Proteins / metabolism
  • Microscopy, Confocal
  • Mutation
  • Oxidation-Reduction
  • Protein Binding
  • Protein Interaction Mapping / methods*
  • Saccharomyces cerevisiae / genetics
  • Saccharomyces cerevisiae / growth & development
  • Saccharomyces cerevisiae / metabolism
  • Substrate Specificity
  • Two-Hybrid System Techniques

Substances

  • Arabidopsis Proteins
  • Luminescent Proteins
  • CDKA1 protein, Arabidopsis
  • Cyclin-Dependent Kinases

Associated data

  • RefSeq/NM_114734
  • RefSeq/NM_179806