Protein expression profiles in Saccharomyces cerevisiae during apoptosis induced by H2O2.
We identified the proteins involved during apoptosis induced by H2O2 in Saccharomyces cerevisiae, and analyzed the global protein pattern by 2-DE. We analyzed classical parameters of apoptosis such as chromatin condensation, DNA fragmentation, and morphology changes of cells. Exposure of yeast cells to nonphysiological doses of peroxides decreases the expression (or increases degradation) of enzymes involved in protection against oxidative stress. This leads the yeast cells to a reduction of their antioxidant defense and makes the cells more prone to apoptosis. In our data the down expression of peroxiredoxin II and GST I, could induce a perturbation of mitochondrial function with an alteration of permeability of the membrane leading to the mitochondria-mediated apoptosis. Moreover, we identified a new spot of a classical glycolytic enzyme: the glyceraldehyde 3-phosphate dehydrogenase during apoptosis. It is known that GAPDH is an extremely abundant glycolytic enzyme with multiple functions and that its overexpression is evident during apoptosis induced by a variety of stimuli. Our results confirm that it is a major intracellular messenger mediating apoptotic death and that this new spot of GAPDH could be an intracellular sensor of oxidative stress during apoptosis induced by H2O2 in S. cerevisiae.
Magherini
Francesca
F
Dipartimento di Scienze Biochimiche, Università degli Studi di Firenze, Firenze, Italy.
Tani
Chiara
C
Gamberi
Tania
T
Caselli
Anna
A
Bianchi
Laura
L
Bini
Luca
L
Modesti
Alessandra
A
D017209:Apoptosis / Q000187:drug effects*; D015153:Blotting, Western; D015180:Electrophoresis, Gel, Two-Dimensional; D020869:Gene Expression Profiling; D024601:Glyceraldehyde 3-Phosphate Dehydrogenase (NADP+) / Q000096:biosynthesis; D006861:Hydrogen Peroxide / Q000494:pharmacology*; D020543:Proteome / Q000737:chemistry; D012441:Saccharomyces cerevisiae / Q000187:drug effects / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000096:biosynthesis*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D020543:Proteome; D029701:Saccharomyces cerevisiae Proteins; D006861:Hydrogen Peroxide; D024601:Glyceraldehyde 3-Phosphate Dehydrogenase (NADP+)