Carbonyl cyanide m-chlorophenylhydrazone induced calcium signaling and activation of plasma membrane H⁺-ATPase in the yeast Saccharomyces cerevisiae

• Published in: FEMS yeast research 2008-06, Vol.8 (4), p.622-630
• Main Authors: Pereira, Michele B.P, Tisi, Renata, Fietto, Luciano G, Cardoso, Anamaria S, França, Mônica M, Carvalho, Fernanda M, Trópia, Maria José M, Martegani, Enzo, Castro, Ieso M, Brandão, Rogelio L
• Format: Article
• Language: English
• Subjects: 2,4-Dinitrophenol; Calcium (extracellular); Calcium (intracellular); Calcium - analysis; Calcium Signaling - drug effects; Calcium signalling; Carbonyl compounds; Carbonyl Cyanide m-Chlorophenyl Hydrazone - analogs & derivatives; Carbonyl Cyanide m-Chlorophenyl Hydrazone - pharmacology; Cell Membrane - enzymology; Cyanides; Cytosol - chemistry; depolarizing compounds; Enzymes; GTP-Binding Protein alpha Subunits - metabolism; H+-transporting ATPase; Hydrogen; Ionophores - pharmacology; Kinases; Phospholipase C; Plasma membrane H+‐ATPase; Protein kinase C; Protein Kinase C - metabolism; Proton-Translocating ATPases - metabolism; Saccharomyces cerevisiae; Saccharomyces cerevisiae - chemistry; Saccharomyces cerevisiae - metabolism; Saccharomyces cerevisiae Proteins - metabolism; Type C Phospholipases - metabolism; Yeast
• ISSN: 1567-1356; 1567-1364
• DOI: 10.1111/j.1567-1364.2008.00380.x

The plasma membrane H⁺-ATPase from Saccharomyces cerevisiae is an enzyme that plays a very important role in the yeast physiology. The addition of protonophores, such as 2,4-dinitrophenol (DNP) and carbonyl cyanide m-chlorophenylhydrazone (CCCP), also triggers a clear in vivo activation of this enzyme. Here, we demonstrate that CCCP-induced activation of the plasma membrane H⁺-ATPase shares some similarities with the sugar-induced activation of the enzyme. Phospholipase C and protein kinase C activities are essential for this activation process while Gpa2p, a G protein involved in the glucose-induced activation of the ATPase, is not required. CCCP also induces a phospholipase C-dependent increase in intracellular calcium. Moreover, we show that the availability of extracellular calcium is required for CCCP stimulation of H⁺-ATPase, suggesting a possible connection between calcium signaling and activation of ATPase.