14-3-3 protects against stress-induced apoptosis.
Expression of human Bax, a cardinal regulator of mitochondrial membrane permeabilization, causes death in yeast. We screened a human cDNA library for suppressors of Bax-mediated yeast death and identified human 14-3-3β/α, a protein whose paralogs have numerous chaperone-like functions. Here, we show that, yeast cells expressing human 14-3-3β/α are able to complement deletion of the endogenous yeast 14-3-3 and confer resistance to a variety of different stresses including cadmium and cycloheximide. The expression of 14-3-3β/α also conferred resistance to death induced by the target of rapamycin inhibitor rapamycin and by starvation for the amino acid leucine, conditions that induce autophagy. Cell death in response to these autophagic stimuli was also observed in the macroautophagic-deficient atg1Δ and atg7Δ mutants. Furthermore, 14-3-3β/α retained its ability to protect against the autophagic stimuli in these autophagic-deficient mutants arguing against so called 'autophagic death'. In line, analysis of cell death markers including the accumulation of reactive oxygen species, membrane integrity and cell surface exposure of phosphatidylserine indicated that 14-3-3β/α serves as a specific inhibitor of apoptosis. Finally, we demonstrate functional conservation of these phenotypes using the yeast homolog of 14-3-3: Bmh1. In sum, cell death in response to multiple stresses can be counteracted by 14-3-3 proteins.
Department of Chemistry and Chemical Engineering, Royal Military College, Kingston, Ontario, Canada.
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Clapp
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Portt
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Khoury
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Sheibani
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Norman
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Ebner
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Eid
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Vali
C A
CA
Mandato
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Madeo
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Greenwood
D048948:14-3-3 Proteins / Q000235:genetics / Q000378:metabolism*; D017209:Apoptosis / Q000187:drug effects; D001343:Autophagy / Q000187:drug effects; D000071193:Autophagy-Related Protein 7; D000071183:Autophagy-Related Proteins; D002104:Cadmium / Q000633:toxicity; D003513:Cycloheximide / Q000633:toxicity; D015723:Gene Library; D006801:Humans; D011494:Protein Kinases / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism; D020123:Sirolimus / Q000494:pharmacology; D051028:bcl-2-Associated X Protein / Q000235:genetics / Q000378:metabolism
D048948:14-3-3 Proteins; C114830:ATG7 protein, S cerevisiae; D000071183:Autophagy-Related Proteins; C075852:BMH1 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; D051028:bcl-2-Associated X Protein; D002104:Cadmium; D003513:Cycloheximide; D011494:Protein Kinases; C477823:ATG1 protein, S cerevisiae; D000071193:Autophagy-Related Protein 7; D020123:Sirolimus