Real-time monitoring of calcineurin activity in living cells: evidence for two distinct Ca2+-dependent pathways in fission yeast

Mol Biol Cell. 2006 Nov;17(11):4790-800. doi: 10.1091/mbc.e06-06-0526. Epub 2006 Aug 23.


In fission yeast, calcineurin dephosphorylates and activates the Prz1 transcription factor. Here, we identified the calcineurin-dependent response element (CDRE) in the promoter region of prz1(+) gene and monitored the calcineurin activity in living cells using a destabilized luciferase reporter gene fused to three tandem repeats of CDRE. Elevated extracellular CaCl(2) caused an increase in calcineurin activity with an initial peak and then approached a sustained constant level in a concentration-dependent manner. In CaCl(2)-sensitive mutants such as Deltapmc1, the response was markedly enhanced, reflecting its high intracellular Ca(2+). Agents expected to induce Ca(2+) influx showed distinct patterns of the CDRE-reporter activity, suggesting different mechanisms of calcineurin activation. Knockout of yam8(+) or cch1(+) encoding putative subunits of a Ca(2+) channel abolished the activation of calcineurin upon exposure to various stimuli, including high extracellular NaCl and cell wall-damaging agents. However, knockout of yam8(+) or cch1(+) did not affect the activation of calcineurin upon stimulation by elevated extracellular Ca(2+). The Pck2 protein kinase C-Pmk1 mitogen-activate protein kinase pathway was required for the stimulation of calcineurin via Yam8/Cch1-mediated Ca(2+) influx, but it was not required for the stimulation by elevated extracellular Ca(2+), suggesting two distinct pathways for calcineurin activation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Calcineurin / genetics
  • Calcineurin / metabolism*
  • Calcium Chloride / pharmacology
  • Calcium Signaling* / drug effects
  • Cell Cycle Proteins / metabolism
  • Chlorpromazine / pharmacology
  • Echinocandins
  • Enzyme Activation / drug effects
  • Enzyme Stability / drug effects
  • Gene Expression
  • Genes, Fungal / genetics
  • Half-Life
  • Lipopeptides
  • Lipoproteins / pharmacology
  • Luciferases / metabolism
  • MAP Kinase Kinase 1 / metabolism
  • Membrane Glycoproteins / metabolism
  • Micafungin
  • Mitogen-Activated Protein Kinases / metabolism
  • Peptides, Cyclic / pharmacology
  • Protein Kinase C / metabolism
  • Response Elements / genetics
  • Schizosaccharomyces / cytology*
  • Schizosaccharomyces / drug effects
  • Schizosaccharomyces / metabolism*
  • Schizosaccharomyces pombe Proteins / metabolism
  • Sodium Chloride / pharmacology


  • Cell Cycle Proteins
  • Echinocandins
  • Lipopeptides
  • Lipoproteins
  • Membrane Glycoproteins
  • Peptides, Cyclic
  • Schizosaccharomyces pombe Proteins
  • YAM8 protein, S pombe
  • Sodium Chloride
  • Luciferases
  • Mek1 protein, S pombe
  • Protein Kinase C
  • Mitogen-Activated Protein Kinases
  • MAP Kinase Kinase 1
  • Calcineurin
  • Calcium Chloride
  • Micafungin
  • Chlorpromazine