Ca2+-Dependent Protein Kinase 6 Enhances KAT2 Shaker Channel Activity in Arabidopsis thaliana

Int J Mol Sci. 2021 Feb 5;22(4):1596. doi: 10.3390/ijms22041596.

Abstract

Post-translational regulations of Shaker-like voltage-gated K+ channels were reported to be essential for rapid responses to environmental stresses in plants. In particular, it has been shown that calcium-dependent protein kinases (CPKs) regulate Shaker channels in plants. Here, the focus was on KAT2, a Shaker channel cloned in the model plant Arabidopsis thaliana, where is it expressed namely in the vascular tissues of leaves. After co-expression of KAT2 with AtCPK6 in Xenopuslaevis oocytes, voltage-clamp recordings demonstrated that AtCPK6 stimulates the activity of KAT2 in a calcium-dependent manner. A physical interaction between these two proteins has also been shown by Förster resonance energy transfer by fluorescence lifetime imaging (FRET-FLIM). Peptide array assays support that AtCPK6 phosphorylates KAT2 at several positions, also in a calcium-dependent manner. Finally, K+ fluorescence imaging in planta suggests that K+ distribution is impaired in kat2 knock-out mutant leaves. We propose that the AtCPK6/KAT2 couple plays a role in the homeostasis of K+ distribution in leaves.

Keywords: Arabidopsis thaliana; CPK; calcium signaling; post-translational regulation; shaker channel.

MeSH terms

  • Animals
  • Arabidopsis / genetics
  • Arabidopsis / metabolism*
  • Arabidopsis Proteins / genetics
  • Arabidopsis Proteins / metabolism*
  • Calcium Signaling
  • Calcium-Calmodulin-Dependent Protein Kinases / genetics
  • Calcium-Calmodulin-Dependent Protein Kinases / metabolism*
  • Female
  • Fluorescence Resonance Energy Transfer
  • Gene Knockout Techniques
  • In Vitro Techniques
  • Models, Molecular
  • Oocytes / metabolism
  • Optical Imaging
  • Patch-Clamp Techniques
  • Phosphorylation
  • Plant Leaves / metabolism
  • Plants, Genetically Modified
  • Potassium / metabolism
  • Potassium Channels, Voltage-Gated / deficiency
  • Potassium Channels, Voltage-Gated / genetics
  • Potassium Channels, Voltage-Gated / metabolism*
  • Recombinant Proteins / genetics
  • Recombinant Proteins / metabolism
  • Xenopus laevis

Substances

  • Arabidopsis Proteins
  • KAT2 protein, Arabidopsis
  • Potassium Channels, Voltage-Gated
  • Recombinant Proteins
  • CPK6 protein, Arabidopsis
  • Calcium-Calmodulin-Dependent Protein Kinases
  • Potassium