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Title:   Author  item
                                            UMER, ANWAR BIN - Rutgers Universityitem
                                            MCLAUGHLIN, JOHN - Rutgers Universityitem
                                            BASU, DEBALEENA - Rutgers Universityitem
                                            McCormick, Susanitem
                                            TUMER, NILGUN - Rutgers University
  Submitted to: Meeting Abstract
                                    Publication Type: Abstract Only
                                    Publication Acceptance Date: 12/9/2010
                                    Publication  Date: 12/9/2010
                                    Citation: Umer, A., Mclaughlin, J., Basu, D., Mccormick, S.P., Tumer, N. 2010. Identification of a direct role for mitochondria in trichothecene resistance. Meeting Abstract.Interpretive Summary:
                                Technical Abstract:   Trichothecenes produced by various species of Fusarium are increasingly contaminating cereal crops worldwide. Fusarium gramineraum causes Fusarium head blight (FHB) in both wheat and barley resulting in reduced plant yield and contamination of the grains with trichothecenes, in particular DON. Improving FHB resistance, hence, remains a high priority in wheat and barley breeding programs throughout the world. Identifying the molecular mechanisms underlying trichothecene toxicity is therefore vital to understanding Fusarium pathology and engineering FHB resistance.
  We have previously shown that mitochondria are critical for trichothecin (Tcin) toxicity in yeast. Sensitivity to Tcin increased when yeast cells were grown in non-fermentable media, which requires functional mitochondria, while cells devoid of mitochondria (p0) showed increased resistance to Tcin. Over 60% of gene deletions that conferred resistance to Tcin were associated with mitochondrial function in our genome wide screening of the yeast deletion library. Moreover, mitochondrial translation was shown to be inhibited by Tcin in the wild type but not in the resistant mutants. To determine if Tcin has a direct effect on mitochondria, we examined translation in isolated yeast mitochondria treated with Tcin. Furthermore, we employed flow cytometry to assess functionality of the yeast mitochondria when treated with trichothecenes. 
  A 60% inhibition in translation was observed in isolated yeast mitochondria treated for 10min with 4μM Tcin, solubilized in 50% ethanol, when compared to mitochondria treated with 50% ethanol. This inhibition increased to 78% at 8μM Tcin suggesting a direct inhibition of mitochondrial translation by Tcin. Flow cytometric analyses of Tcin-treated yeast cells stained for mitochondrial membrane potential, ROS generation and cell death also suggest a role for mitochondria in Tcin-induced cell death. Peak shifts in the median fluorescence intensities of Tcin-treated cells indicate that Tcin triggers ROS generation resulting in hyperpolarization of the mitochondrial membrane which eventually leads to cell death.


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