title: Activation of a TRP-like channel and intracellular Ca2+ dynamics during phospholipase-C-mediated cell death.
abstract: The model organism Neurospora crassa undergoes programmed cell death when exposed to staurosporine. Here, we show that staurosporine causes defined changes in cytosolic free Ca(2+) ([Ca(2+)]c) dynamics and a distinct Ca(2+) signature that involves Ca(2+) influx from the external medium and internal Ca(2+) stores. We investigated the molecular basis of this Ca(2+) response by using [Ca(2+)]c measurements combined with pharmacological and genetic approaches. Phospholipase C was identified as a pivotal player during cell death, because modulation of the phospholipase C signaling pathway and deletion of PLC-2, which we show to be involved in hyphal development, results in an inability to trigger the characteristic staurosporine-induced Ca(2+) signature. Using Δcch-1, Δfig-1 and Δyvc-1 mutants and a range of inhibitors, we show that extracellular Ca(2+) entry does not occur through the hitherto described high- and low-affinity Ca(2+) uptake systems, but through the opening of plasma membrane channels with properties resembling the transient receptor potential (TRP) family. Partial blockage of the response to staurosporine after inhibition of a putative inositol-1,4,5-trisphosphate (IP3) receptor suggests that Ca(2+) release from internal stores following IP3 formation combines with the extracellular Ca(2+) influx.
authors.affiliation: IBMC-Instituto de Biologia Molecular e Celular - Universidade do Porto, Rua do Campo Alegre 823, 4150-180 Porto, Portugal ICBAS-Instituto de Ciências Biomédicas de Abel Salazar, Universidade do Porto, Rua de Jorge Viterbo Ferreira 228, 4050-313 Porto, Portugal apgoncalves@ibmc.up.pt avideira@ibmc.up.pt.
authors.forename: A Pedro
authors.initials: AP
authors.lastname: Gonçalves
authors.affiliation: ICBAS-Instituto de Ciências Biomédicas de Abel Salazar, Universidade do Porto, Rua de Jorge Viterbo Ferreira 228, 4050-313 Porto, Portugal.
authors.forename: J Miguel
authors.initials: JM
authors.lastname: Cordeiro
authors.affiliation: IBMC-Instituto de Biologia Molecular e Celular - Universidade do Porto, Rua do Campo Alegre 823, 4150-180 Porto, Portugal.
authors.forename: João
authors.initials: J
authors.lastname: Monteiro
authors.affiliation: Manchester Fungal Infection Group, Institute of Inflammation and Repair, CTF Building, Grafton Street, University of Manchester, Manchester M13 9NT, UK.
authors.forename: Alberto
authors.initials: A
authors.lastname: Muñoz
authors.affiliation: ICBAS-Instituto de Ciências Biomédicas de Abel Salazar, Universidade do Porto, Rua de Jorge Viterbo Ferreira 228, 4050-313 Porto, Portugal.
authors.forename: Paulo
authors.initials: P
authors.lastname: Correia-de-Sá
authors.affiliation: Manchester Fungal Infection Group, Institute of Inflammation and Repair, CTF Building, Grafton Street, University of Manchester, Manchester M13 9NT, UK.
authors.forename: Nick D
authors.initials: ND
authors.lastname: Read
authors.affiliation: IBMC-Instituto de Biologia Molecular e Celular - Universidade do Porto, Rua do Campo Alegre 823, 4150-180 Porto, Portugal ICBAS-Instituto de Ciências Biomédicas de Abel Salazar, Universidade do Porto, Rua de Jorge Viterbo Ferreira 228, 4050-313 Porto, Portugal apgoncalves@ibmc.up.pt avideira@ibmc.up.pt.
authors.forename: Arnaldo
authors.initials: A
authors.lastname: Videira
mesh_terms: D000818:Animals; D002118:Calcium / Q000378:metabolism*; D015220:Calcium Channels / Q000378:metabolism; D016923:Cell Death; D002462:Cell Membrane / Q000378:metabolism; D003593:Cytoplasm / Q000378:metabolism*; D015544:Inositol 1,4,5-Trisphosphate / Q000378:metabolism*; D009492:Neurospora crassa; D015398:Signal Transduction / Q000502:physiology; D010738:Type C Phospholipases / Q000378:metabolism*
chemical_list: D015220:Calcium Channels; D015544:Inositol 1,4,5-Trisphosphate; D010738:Type C Phospholipases; D002118:Calcium
keywords: Cell death; Inositol-1,4,5-trisphosphate; Phospholipase C; Staurosporine; Transient receptor potential channel