title: Yeast growth in raffinose results in resistance to acetic-acid induced programmed cell death mostly due to the activation of the mitochondrial retrograde pathway.
abstract: In order to investigate whether and how a modification of mitochondrial metabolism can affect yeast sensitivity to programmed cell death (PCD) induced by acetic acid (AA-PCD), yeast cells were grown on raffinose, as a sole carbon source, which, differently from glucose, favours mitochondrial respiration. We found that, differently from glucose-grown cells, raffinose-grown cells were mostly resistant to AA-PCD and that this was due to the activation of mitochondrial retrograde (RTG) response, which increased with time, as revealed by the up-regulation of the peroxisomal isoform of citrate synthase and isocitrate dehydrogenase isoform 1, RTG pathway target genes. Accordingly, the deletion of RTG2 and RTG3, a positive regulator and a transcription factor of the RTG pathway, resulted in AA-PCD, as shown by TUNEL assay. Neither deletion in raffinose-grown cells of HAP4, encoding the positive regulatory subunit of the Hap2,3,4,5 complex nor constitutive activation of the RTG pathway in glucose-grown cells due to deletion of MKS1, a negative regulator of RTG pathway, had effect on yeast AA-PCD. The RTG pathway was found to be activated in yeast cells containing mitochondria, in which membrane potential was measured, capable to consume oxygen in a manner stimulated by the uncoupler CCCP and inhibited by the respiratory chain inhibitor antimycin A. AA-PCD resistance in raffinose-grown cells occurs with a decrease in both ROS production and cytochrome c release as compared to glucose-grown cells en route to AA-PCD.
authors.affiliation: CNR, Istituto di Biomembrane e Bioenergetica, Via Amendola 165/a, 70126 Bari, Italy.
authors.forename: Nicoletta
authors.initials: N
authors.lastname: Guaragnella
authors.affiliation: CNR, Istituto di Biomembrane e Bioenergetica, Via Amendola 165/a, 70126 Bari, Italy.
authors.forename: Maša
authors.initials: M
authors.lastname: Ždralević
authors.affiliation: CNR, Istituto di Biomembrane e Bioenergetica, Via Amendola 165/a, 70126 Bari, Italy.
authors.forename: Paolo
authors.initials: P
authors.lastname: Lattanzio
authors.affiliation: CNR, Istituto di Biomembrane e Bioenergetica, Via Amendola 165/a, 70126 Bari, Italy.
authors.forename: Domenico
authors.initials: D
authors.lastname: Marzulli
authors.affiliation: Department of Biological Sciences, University of New Orleans, 2000 Lakeshore Drive, New Orleans, LA 70148, USA.
authors.forename: Tammy
authors.initials: T
authors.lastname: Pracheil
authors.affiliation: Department of Biological Sciences, University of New Orleans, 2000 Lakeshore Drive, New Orleans, LA 70148, USA.
authors.forename: Zhengchang
authors.initials: Z
authors.lastname: Liu
authors.affiliation: Dipartimento di Medicina e Scienze per la Salute, Università del Molise, Via de Sanctis, 86100 Campobasso, Italy.
authors.forename: Salvatore
authors.initials: S
authors.lastname: Passarella
authors.affiliation: CNR, Istituto di Biomembrane e Bioenergetica, Via Amendola 165/a, 70126 Bari, Italy.
authors.forename: Ersilia
authors.initials: E
authors.lastname: Marra
authors.affiliation: CNR, Istituto di Biomembrane e Bioenergetica, Via Amendola 165/a, 70126 Bari, Italy. Electronic address: s.giannattasio@ibbe.cnr.it.
authors.forename: Sergio
authors.initials: S
authors.lastname: Giannattasio
mesh_terms: D019342:Acetic Acid / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects*; D045304:Cytochromes c / Q000378:metabolism; D017353:Gene Deletion; D005947:Glucose / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration / Q000187:drug effects; D015151:Immunoblotting; D042541:Intracellular Space / Q000187:drug effects / Q000378:metabolism; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects; D008928:Mitochondria / Q000187:drug effects / Q000378:metabolism*; D010766:Phosphorylation / Q000187:drug effects; D011887:Raffinose / Q000494:pharmacology*; D017382:Reactive Oxygen Species / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000166:cytology / Q000187:drug effects / Q000254:growth & development*; D029701:Saccharomyces cerevisiae Proteins / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects*
chemical_list: D017382:Reactive Oxygen Species; D029701:Saccharomyces cerevisiae Proteins; D045304:Cytochromes c; D005947:Glucose; D011887:Raffinose; D019342:Acetic Acid
keywords: 2,7-dihydrodichlorofluorescein diacetate; AA; Acetic acid; CCCP; DCF; GLU; H(2)DCH-DA; Mitochondria; N-acetyl cysteine; NAC; PCD; Programmed cell death; RAF; RTG; Raffinose; Retrograde pathway; TMRM; TUNEL; WT; Yeast; acetic acid; carbonyl cyanide m-chlorophenylhydrazone; cyt c; cytochrome c; dichlorophluoresceine; glucose-grown; programmed cell death; raffinose-grown; retrograde; terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick end labelling; tetramethylrhodamine methyl ester; wild type