Journal: Nature communications

Article Title: Superoxide dismutase 1 acts as a nuclear transcription factor to regulate oxidative stress resistance

doi: 10.1038/ncomms4446

Figure Lengend Snippet: (a) Sod1 is required for the induction of oxidative response (OR) genes. WT or sod1 Δ cells were treated without or with 0.4 mM H 2 O 2 for 20 min and analyzed for global gene expression profile. 123 Sod1-dependent genes were identified and most of the known genes belong to five related functional categories. (b) Shown is the relative induction level of OR genes by H 2 O 2 in each category in WT and sod1 Δ cells. Data represents average fold change of induction in each category. (c) Shown is the heat map of genes in the oxidative stress response category. (d) Validation of representative genes ( GRE2 , Genes de Respuesta a Estres 2; TSA2 : Thiol-Specific Antioxidant 2; YML131; STF2 : STabilizing Factor 2) in the oxidative stress response category by RT-qPCR. Error bars indicate ± SD from triplicates of two independent experiments. * p < 0.05. (e) Nuclear Sod1 is critical for the induction of OR genes. Yeast cells expressing different forms of Sod1 were treated with 0.4 mM H 2 O 2 for 20 min. Representative genes were validated by RT-qPCR. Error bars indicate ± SD from triplicates of two independent experiments. * p < 0.05. (f) The induction of OR genes by ROS was attenuated in Sod1 S60,99A cells. Yeast cells expressing Sod1 or Sod1 S60,99A were treated with 0.4 mM H 2 O 2 for 20 min. Expression of GRE2 and RNR3 were determined by RT-qPCR. Error bars indicate ± SD from triplicates of two independent experiments. * p < 0.05. (g) ROS treatment increases the association of Sod1 with promoter of oxidative responsive genes. WT (SZy1051) and sod1 Δ (SZy1050) cells were treated with 0.4 mM H 2 O 2 for 20 min. The binding of Sod1 to representative promoters were analyzed by chromatin immunoprecipitation (ChIP). (h) Quantification of the Fig. 5g experiment. Error bars indicate ± SD from triplicates of two independent experiments. * p < 0.05, Student’s t -test.

Article Snippet: DNA microarray analysis of yeast global gene expression was carried out by Rutgers RUCDR Analytical and Informatics Services using the following procedure.

Techniques: Gene Expression, Functional Assay, Biomarker Discovery, Quantitative RT-PCR, Expressing, Binding Assay, Chromatin Immunoprecipitation