Disruption of genes encoding pyruvate dehydrogenase kinases leads to retarded growth on acetate and ethanol in <i>Saccharomyces cerevisiae</i>

Steensma, H. Yde; Tomáška, Ľubomír; Reuven, Peter; Nosek, Jozef; Brandt, Raymond

doi:10.1002/yea.1543

Cited by 10 publications

(15 citation statements)

References 33 publications

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“…4C) may suggest that the 2 sites represent a combinatorial output, not a strictly hierarchical one whereby position 240 affects the phosphorylation state of threonine 199. The 2 known yeast mitochondrial kinases (Pkp1 and Pkp2) have been identified as orthologs of mammalian PDKs (pyruvate dehydrogenase kinases), which exist as a family of paralogs with different tissue distribution in metazoans (44). Reports in the literature (45,46) also link the 2 phosphatases, Ptc5 and Aup1 (Ptc6), to Pyruvate Dehydrogenase Alpha subunit (Pda1) dephosphorylation in yeast.…”

Section: Discussionmentioning

confidence: 99%

Phosphorylation of mitochondrial matrix proteins regulates their selective mitophagic degradation

Kolitsida

Zhou

Rackiewicz

et al. 2019

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

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Mitophagy is an important quality-control mechanism in eukaryotic cells, and defects in mitophagy correlate with aging phenomena and neurodegenerative disorders. It is known that different mitochondrial matrix proteins undergo mitophagy with very different rates but, to date, the mechanism underlying this selectivity at the individual protein level has remained obscure. We now present evidence indicating that protein phosphorylation within the mitochondrial matrix plays a mechanistic role in regulating selective mitophagic degradation in yeast via involvement of the Aup1 mitochondrial protein phosphatase, as well as 2 known matrix-localized protein kinases, Pkp1 and Pkp2. By focusing on a specific matrix phosphoprotein reporter, we also demonstrate that phospho-mimetic and nonphosphorylatable point mutations at known phosphosites in the reporter increased or decreased its tendency to undergo mitophagy. Finally, we show that phosphorylation of the reporter protein is dynamically regulated during mitophagy in an Aup1-dependent manner. Our results indicate that structural determinants on a mitochondrial matrix protein can govern its mitophagic fate, and that protein phosphorylation regulates these determinants.

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Section: Discussionmentioning

confidence: 99%

Phosphorylation of mitochondrial matrix proteins regulates their selective mitophagic degradation

Kolitsida

Zhou

Rackiewicz

et al. 2019

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

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“…4C). The mitochondrial origin of these structures was confirmed by co-localization of the Mitotracker-positive sheets with fluorescently tagged Pda1p, the E1 alpha subunit of the pyruvate dehydrogenase complex that concentrates in spots within the mitochondrial matrix [49,50]. Interestingly, in contrast to the tubular mitochondria, where Pda1-GFP signal covered most of the Mitotracker-stained volume at the resolution of fluorescence microscopy, Pda1-GFP spots overlapping with Mitotracker-stained sheets were confined exclusively to the border of these sheets (Fig.…”

Section: Resultsmentioning

confidence: 99%

“…In addition, our co-localization experiments revealed a specific structure of the sheets. The mitochondrial matrix protein, Pda1p [49,50], localized exclusively to distinct foci at the border of the sheets, while the inner area of the sheet remained void of Pda1p-GFP signal, suggesting fragmentation of the IMM within these altered mitochondria.…”

Section: Discussionmentioning

confidence: 99%

Specific degradation of phosphatidylglycerol is necessary for proper mitochondrial morphology and function

Pokorná

C̆ermáková

Horváth

et al. 2016

Biochimica et Biophysica Acta (BBA) - Bioenergetics

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In yeast, phosphatidylglycerol (PG) is a minor phospholipid under standard conditions; it can be utilized for cardiolipin (CL) biosynthesis by CL synthase, Crd1p, or alternatively degraded by the phospholipase Pgc1p. The Saccharomyces cerevisiae deletion mutants crd1Δ and pgc1Δ both accumulate PG. Based on analyses of the phospholipid content of pgc1Δ and crd1Δ yeast, we revealed that in yeast mitochondria, two separate pools of PG are present, which differ in their fatty acid composition and accessibility for Pgc1p-catalyzed degradation. In contrast to CL-deficient crd1Δ yeast, the pgc1Δ mutant contains normal levels of CL. This makes the pgc1Δ strain a suitable model to study the effect of accumulation of PG per se. Using fluorescence microscopy, we show that accumulation of PG with normal levels of CL resulted in increased fragmentation of mitochondria, while in the absence of CL, accumulation of PG led to the formation of large mitochondrial sheets. We also show that pgc1Δ mitochondria exhibited increased respiration rates due to increased activity of cytochrome c oxidase. Taken together, our results indicate that not only a lack of anionic phospholipids, but also excess PG, or unbalanced ratios of anionic phospholipids in mitochondrial membranes, have harmful consequences on mitochondrial morphology and function.

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“…Disruption of PTC5 gene (ORF YGL059 ) had little effect on yeast growth on glucose, but caused retardation on acetate. Growth rate of the mutant was restored to the wild-type level by simultaneous disruption of the PDA1 gene, which encodes the α subunit of pyruvate dehydrogenase complex [23]. A further study showed that both Ptc5 and Ptc6 ( YCR079 ) are involved in the regulation of this complex by affecting the phosphorylation state of Pda1 [24].…”

Section: Introductionmentioning

confidence: 97%

A Type 2C Protein Phosphatase FgPtc3 Is Involved in Cell Wall Integrity, Lipid Metabolism, and Virulence in Fusarium graminearum

et al. 2011

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Type 2C protein phosphatases (PP2Cs) play important roles in regulating many biological processes in eukaryotes. Currently, little is known about functions of PP2Cs in filamentous fungi. The causal agent of wheat head blight, Fusarium graminearum, contains seven putative PP2C genes, FgPTC1, -3, -5, -5R, -6, -7 and -7R. In order to investigate roles of these PP2Cs, we constructed deletion mutants for all seven PP2C genes in this study. The FgPTC3 deletion mutant (ΔFgPtc3-8) exhibited reduced aerial hyphae formation and deoxynivalenol (DON) production, but increased production of conidia. The mutant showed increased resistance to osmotic stress and cell wall-damaging agents on potato dextrose agar plates. Pathogencity assays showed that ΔFgPtc3-8 is unable to infect flowering wheat head. All of the defects were restored when ΔFgPtc3-8 was complemented with the wild-type FgPTC3 gene. Additionally, the FgPTC3 partially rescued growth defect of a yeast PTC1 deletion mutant under various stress conditions. Ultrastructural and histochemical analyses showed that conidia of ΔFgPtc3-8 contained an unusually high number of large lipid droplets. Furthermore, the mutant accumulated a higher basal level of glycerol than the wild-type progenitor. Quantitative real-time PCR assays showed that basal expression of FgOS2, FgSLT2 and FgMKK1 in the mutant was significantly higher than that in the wild-type strain. Serial analysis of gene expression in ΔFgPtc3-8 revealed that FgPTC3 is associated with various metabolic pathways. In contrast to the FgPTC3 mutant, the deletion mutants of FgPTC1, FgPTC5, FgPTC5R, FgPTC6, FgPTC7 or FgPTC7R did not show aberrant phenotypic features when grown on PDA medium or inoculated on wheat head. These results indicate FgPtc3 is the key PP2C that plays a critical role in a variety of cellular and biological functions, including cell wall integrity, lipid and secondary metabolisms, and virulence in F. graminearum.

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Disruption of genes encoding pyruvate dehydrogenase kinases leads to retarded growth on acetate and ethanol in Saccharomyces cerevisiae

Cited by 10 publications

References 33 publications

Phosphorylation of mitochondrial matrix proteins regulates their selective mitophagic degradation

Phosphorylation of mitochondrial matrix proteins regulates their selective mitophagic degradation

Specific degradation of phosphatidylglycerol is necessary for proper mitochondrial morphology and function

A Type 2C Protein Phosphatase FgPtc3 Is Involved in Cell Wall Integrity, Lipid Metabolism, and Virulence in Fusarium graminearum

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