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

Jiang, Heng; English, Ann M.

doi:10.1016/j.jinorgbio.2006.07.017

Cited by 30 publications

(16 citation statements)

References 60 publications

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“…These were prepared as previously described [30]. Cells were collected at 2000 g , washed twice with 20 mM potassium phosphate buffer (KPi, pH 7.0) containing 0.1 mM PMSF, the pellets were suspended in the same buffer ±0.1 M iodoacetamide, and mixed with an equal volume of acid-washed glass beads.…”

Section: Methodsmentioning

confidence: 99%

SOD1 oxidation and formation of soluble aggregates in yeast: Relevance to sporadic ALS development

Martins¹,

English²

2014

Redox Biology

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Misfolding and aggregation of copper–zinc superoxide dismutase (Sod1) are observed in neurodegenerative diseases such as amyotrophic lateral sclerosis (ALS). Mutations in Sod1 lead to familial ALS (FALS), which is a late-onset disease. Since oxidative damage to proteins increases with age, it had been proposed that oxidation of Sod1 mutants may trigger their misfolding and aggregation in FALS. However, over 90% of ALS cases are sporadic (SALS) with no obvious genetic component. We hypothesized that oxidation could also trigger the misfolding and aggregation of wild-type Sod1 and sought to confirm this in a cellular environment. Using quiescent, stationary-phase yeast cells as a model for non-dividing motor neurons, we probed for post-translational modification (PTM) and aggregation of wild-type Sod1 extracted from these cells. By size-exclusion chromatography (SEC), we isolated two populations of Sod1 from yeast: a low-molecular weight (LMW) fraction that is catalytically active and a catalytically inactive, high-molecular weight (HMW) fraction. High-resolution mass spectrometric analysis revealed that LMW Sod1 displays no PTMs but HMW Sod1 is oxidized at Cys146 and His71, two critical residues for the stability and folding of the enzyme. HMW Sod1 is also oxidized at His120, a copper ligand, which will promote loss of this catalytic metal cofactor essential for SOD activity. Monitoring the fluorescence of a Sod1-green-fluorescent-protein fusion (Sod1-GFP) extracted from yeast chromosomally expressing this fusion, we find that HMW Sod1-GFP levels increase up to 40-fold in old cells. Thus, we speculate that increased misfolding and inclusion into soluble aggregates is a consequence of elevated oxidative modifications of wild-type Sod1 as cells age. Our observations argue that oxidative damage to wild-type Sod1 initiates the protein misfolding mechanisms that give rise to SALS.

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Section: Methodsmentioning

confidence: 99%

SOD1 oxidation and formation of soluble aggregates in yeast: Relevance to sporadic ALS development

Martins¹,

English²

2014

Redox Biology

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“…functions (9)(10)(11), Ccp1 may serve in future as a paradigm of hemebased H 2 O 2 sensing and heme transfer.…”

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confidence: 99%

Respiration triggers heme transfer from cytochrome c peroxidase to catalase in yeast mitochondria

Kathiresan

Martins

English

2014

Proc. Natl. Acad. Sci. U.S.A.

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Significance We provide to our knowledge the first in vivo and in vitro evidence for H 2 O 2 -triggered heme transfer between proteins. Specifically, H 2 O 2 binds to and labilizes cytochrome c peroxidase (Ccp1)’s heme by oxidizing the proximal Fe ligand (His175), which activates Ccp1 to transfer its heme to apoCta1, and apoCcp1 subsequently escapes from mitochondria. This sequence of H 2 O 2 -activated heme labilization, heme transfer between proteins, and protein relocalization defines a previously undefined mechanism of H 2 O 2 signaling in cells. In contrast, established H 2 O 2 signaling mechanisms are dominated by thiol-based redox changes.

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“…Protein extracts from stressed and control cultures were prepared as reported [23,30]. Briefly, cells were washed twice with 100 mM potassium phosphate buffer, pH 7.0 (KP i ) and centrifuged at 2000 g, the pellets were suspended in KP i with 0.1 mM phenylmethanesulfonyl fluoride and mixed with an equal volume of acid-washed glass beads.…”

Section: Preparation Of Soluble Protein Extracts and Enzyme Activity mentioning

confidence: 99%

“…In fact, the production of Ccp1 [28] and of a lacZ reporter controlled by the Ccp1 promoter [29] increases on H 2 O 2 challenge, and Ccp1 deletion heightens the sensitivity of yeast to exogenous H 2 O 2 [23,[29][30][31]. Nonetheless, whereas ccp1Δ cells are hypersensitive to H 2 O 2 [23], cells expressing the Ccp1 W191F variant with negligible Cyc II -oxidizing activity [32] are more resistant to H 2 O 2 challenge than wild-type cells [23,29].…”

Section: Introductionmentioning

confidence: 99%

“…Nonetheless, whereas ccp1Δ cells are hypersensitive to H 2 O 2 [23], cells expressing the Ccp1 W191F variant with negligible Cyc II -oxidizing activity [32] are more resistant to H 2 O 2 challenge than wild-type cells [23,29]. This led us [23,30] and others [33] to identify Ccp1 as an H 2 O 2 -sensing and signaling molecule. H 2 O 2 exposure of Ccp1-and Ccp1 W191F -producing strains increases total peroxiredoxin (Prx) and catalase activities [23] as well as expression of the mRNA for Tsa1, the major yeast Prx [33].…”

Section: Introductionmentioning

confidence: 99%

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Peroxynitrite and hydrogen peroxide elicit similar cellular stress responses mediated by the Ccp1 sensor protein

Martins

Bakas

McIntosh

et al. 2015

Free Radical Biology and Medicine

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

Cited by 30 publications

References 60 publications

SOD1 oxidation and formation of soluble aggregates in yeast: Relevance to sporadic ALS development

SOD1 oxidation and formation of soluble aggregates in yeast: Relevance to sporadic ALS development

Respiration triggers heme transfer from cytochrome c peroxidase to catalase in yeast mitochondria

Peroxynitrite and hydrogen peroxide elicit similar cellular stress responses mediated by the Ccp1 sensor protein

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