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First published online 28 August 2007
doi: 10.1242/jcs.010926

Journal of Cell Science 120, 3279-3288 (2007)
Published by The Company of Biologists 2007
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Research Article

NO-mediated apoptosis in yeast

Bruno Almeida1, Sabrina Buttner2, Steffen Ohlmeier3, Alexandra Silva1, Ana Mesquita1, Belém Sampaio-Marques1, Nuno S. Osório1, Alexander Kollau4, Bernhard Mayer4, Cecília Leão1, João Laranjinha5, Fernando Rodrigues1, Frank Madeo2 and Paula Ludovico1,*

1 Life and Health Sciences Research Institute (ICVS), School of Health Sciences, University of Minho, Campus de Gualtar, 4710-057 Braga, Portugal
2 Institute for Molecular Biosciences, Universitätsplatz 2, A-8010 Graz, Austria
3 Proteomics Core Facility, Biocenter Oulu, Department of Biochemistry, University of Oulu, Oulu, Finland
4 Department of Pharmacology and Toxicology, KFUG, Universitätsplatz 2, A-8010 Graz, Austria
5 Faculty of Pharmacy and Center for Neurosciences and Cell Biology, University of Coimbra, 3000 Coimbra, Portugal

* Author for correspondence (e-mail: pludovico{at}ecsaude.uminho.pt)

Accepted 18 July 2007

Nitric oxide (NO) is a small molecule with distinct roles in diverse physiological functions in biological systems, among them the control of the apoptotic signalling cascade. By combining proteomic, genetic and biochemical approaches we demonstrate that NO and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) are crucial mediators of yeast apoptosis. Using indirect methodologies and a NO-selective electrode, we present results showing that H2O2-induced apoptotic cells synthesize NO that is associated to a nitric oxide synthase (NOS)-like activity as demonstrated by the use of a classical NOS kit assay. Additionally, our results show that yeast GAPDH is a target of extensive proteolysis upon H2O2-induced apoptosis and undergoes S-nitrosation. Blockage of NO synthesis with N{omega}-nitro-L-arginine methyl ester leads to a decrease of GAPDH S-nitrosation and of intracellular reactive oxygen species (ROS) accumulation, increasing survival. These results indicate that NO signalling and GAPDH S-nitrosation are linked with H2O2-induced apoptotic cell death. Evidence is presented showing that NO and GAPDH S-nitrosation also mediate cell death during chronological life span pointing to a physiological role of NO in yeast apoptosis.

Key words: Yeast apoptosis, Nitric oxide, S-nitrosation, Glyceraldehyde-3-phosphate dehydrogenase, L-arginine, Reactive oxygen species


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