Phenotypic analysis of the ccp1Δ and ccp1Δ-ccp1W191F mutant strains of Saccharomyces cerevisiae indicates that cytochrome c peroxidase functions in oxidative-stress signaling

Yeast cytochrome c peroxidase (CCP) efficiently catalyzes the reduction of H2O2 to H2O by ferrocytochrome c in vitro. The physiological function of CCP, a heme peroxidase that is targeted to the mitochondrial intermembrane space of Saccharomyces cereuisiae, is not known. CCP1-null-mutant cells in the W303-1B genetic background (ccp1 Delta) grew as well as wild-type cells with glucose, ethanol, glycerol or lactate as carbon sources but with a shorter initial doubling time. Monitoring growth over 10 days demonstrated that CCP1 does not enhance mitochondrial function in unstressed cells. No role for CCP1 was apparent in cells exposed to heat stress under aerobic or anaerobic conditions. However, the detoxification function of CCP protected respiring mitochondria when cells were challenged with H2O2. Transformation of ccp1 Delta with ccp1(W191F) which encodes the CCPW191F mutant enzyme lacking CCP activity, significantly increased the sensitivity to H2O2 of exponential-phase fermenting cells. In contrast, stationary-phase (7-day) ccp1 Delta-ccp1(W191F) exhibited wild-type tolerance to H2O2, which exceeded that of ccp1 Delta. Challenge with H2O2 caused increased CCP, superoxide dismutase and catalase antioxidant enzyme activities (but not glutathione reductase activity) in exponentially growing cells and decreased antioxidant activities in stationary-phase cells. Although unstressed stationary-phase ccp1 Delta exhibited the highest catalase and glutathione reductase activities, a greater loss of these antioxidant activities was observed on H2O2 exposure in ccp1 Delta than in ccp1 Delta-ccp1(W191F) and wild-type cells. The phenotypic differences reported here between the ccp1 Delta and ccp1 Delta-ccp1(W191F) strains lacking CCP activity provide strong evidence that CCP has separate antioxidant and signaling functions in yeast. (c) 2006 Elsevier Inc. All rights reserved.