RRC ID 15296
著者 Ma Y, Sugiura R, Koike A, Ebina H, Sio SO, Kuno T.
タイトル Transient receptor potential (TRP) and Cch1-Yam8 channels play key roles in the regulation of cytoplasmic Ca2+ in fission yeast.
ジャーナル PLoS One
Abstract The regulation of cytoplasmic Ca(2+) is crucial for various cellular processes. Here, we examined the cytoplasmic Ca(2+) levels in living fission yeast cells by a highly sensitive bioluminescence resonance energy transfer-based assay using GFP-aequorin fusion protein linked by 19 amino acid. We monitored the cytoplasmic Ca(2+) level and its change caused by extracellular stimulants such as CaCl(2) or NaCl plus FK506 (calcineurin inhibitor). We found that the extracellularly added Ca(2+) caused a dose-dependent increase in the cytoplasmic Ca(2+) level and resulted in a burst-like peak. The overexpression of two transient receptor potential (TRP) channel homologues, Trp1322 or Pkd2, markedly enhanced this response. Interestingly, the burst-like peak upon TRP overexpression was completely abolished by gene deletion of calcineurin and was dramatically decreased by gene deletion of Prz1, a downstream transcription factor activated by calcineurin. Furthermore, 1 hour treatment with FK506 failed to suppress the burst-like peak. These results suggest that the burst-like Ca(2+) peak is dependent on the transcriptional activity of Prz1, but not on the direct TRP dephosphorylation. We also found that extracellularly added NaCl plus FK506 caused a synergistic cytosolic Ca(2+) increase that is dependent on the inhibition of calcineurin activity, but not on the inhibition of Prz1. The synergistic Ca(2+) increase is abolished by the addition of the Ca(2+) chelator BAPTA into the media, and is also abolished by deletion of the gene encoding a subunit of the Cch1-Yam8 Ca(2+) channel complex, indicating that the synergistic increase is caused by the Ca(2+) influx from the extracellular medium via the Cch1-Yam8 complex. Furthermore, deletion of Pmk1 MAPK abolished the Ca(2+) influx, and overexpression of the constitutively active Pek1 MAPKK enhanced the influx. These results suggest that Pmk1 MAPK and calcineurin positively and negatively regulate the Cch1-Yam8 complex, respectively, via modulating the balance between phosphorylation and dyphosphorylation state.
巻・号 6(7)
ページ e22421
公開日 2011-1-1
DOI 10.1371/journal.pone.0022421
PII PONE-D-11-04862
PMID 21811607
PMC PMC3139647
MeSH Aequorin / metabolism Calcineurin / metabolism Calcineurin Inhibitors Calcium / metabolism* Calcium Channels / metabolism* Calcium Chloride / pharmacology Cell Membrane Permeability / drug effects Cytoplasm / drug effects Cytoplasm / metabolism* Extracellular Space / drug effects Extracellular Space / metabolism Gene Knockout Techniques Genes, Fungal / genetics Golgi Apparatus / drug effects Golgi Apparatus / metabolism Green Fluorescent Proteins / metabolism Immunoblotting Membrane Glycoproteins / metabolism* Microscopy, Fluorescence Recombinant Fusion Proteins / metabolism Schizosaccharomyces / cytology Schizosaccharomyces / drug effects Schizosaccharomyces / genetics Schizosaccharomyces / metabolism* Schizosaccharomyces pombe Proteins / metabolism* Sodium Chloride / pharmacology Tacrolimus / pharmacology Transcription, Genetic / drug effects Transient Receptor Potential Channels / metabolism*
IF 2.74
引用数 25
WOS 分野 BIOCHEMISTRY & MOLECULAR BIOLOGY
リソース情報
酵母 FY17210(KP2101)