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MBC in Press, published online ahead of print August 9, 2006
Mol. Biol. Cell 10.1091/mbc.E06-05-0475

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Submitted on May 31, 2006
Revised on July 28, 2006
Accepted on August 1, 2006

Characterization of DNA Damage in Yeast Apoptosis Induced by Hydrogen Peroxide, Acetic Acid, and Hyperosmotic Shock

Gabriela F. Ribeiro, Manuela Côrte-Real, and Björn Johansson

Departamento de Biologia-Centro de Biologia Universidade do Minho, Campus de Gualtar, 4710-057 Braga, Portugal

Monitoring Editor: Thomas Fox

S. cerevisiae has been reported to die, under certain conditions, from programmed cell death with apoptotic markers. One of the most important markers is chromosomal DNA fragmentation as indicated by TUNEL staining. We found TUNEL staining in S. cerevisiae to be a consequence of both single and double strand DNA breaks while In situ ligation specifically stained double strand DNA breaks. Cells treated with hydrogen peroxide or acetic acid staining positively for TUNEL assay stained negatively for In situ ligation, indicating that DNA damage in both cases mainly consists of single strand DNA breaks. Pulsed Field Gel Electrophoresis of chromosomal DNA from cells dying from hydrogen peroxide, acetic acid or hyperosmotic shock, revealed DNA breakdown into fragments of several hundred kilobases, consistent with the higher order chromatin degradation preceding DNA laddering in apoptotic mammalian cells. DNA fragmentation was associated with death by treatment with 10 mM hydrogen peroxide but not 150 mM and was absent if cells were fixed with formaldehyde to eliminate enzyme activity before hydrogen peroxide treatment. These observations are consistent with a process that, like mammalian apoptosis, is enzyme dependent, degrades chromosomal DNA, and is activated only at low intensity of death stimuli.

Address correspondence to: Björn Johansson (bjorn_johansson{at}bio.uminho.pt)




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 Nucleic Acids ResHome page
S. Mroczek and J. Kufel
Apoptotic signals induce specific degradation of ribosomal RNA in yeast
Nucleic Acids Res., May 1, 2008; 36(9): 2874 - 2888.
[Abstract] [Full Text] [PDF]


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