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| Status |
Public on Mar 20, 2023 |
| Title |
Temporal Coordination of the Transcription Factor Response to H2O2stress [bulk RNA-seq] |
| Organism |
Homo sapiens |
| Experiment type |
Expression profiling by high throughput sequencing
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| Summary |
The p53 and FOXO transcription factors (TFs) share many similarities despite their distinct evolutionary origins. Both TFs are activated by a variety of cellular stresses and upregulate genes in similar pathways including cell-cycle arrest and apoptosis. Oxidative stress from excess H2O2activates both FOXO1 and p53, yet whether they are activated at the same time is unclear. Here we found that cells respond to high H2O2levels in two temporal phases. In the first phase FOXO1 rapidly shuttles to the nucleus while p53 levels remain low. In the second phase FOXO1 exits the nucleus and p53 levels rise. We found that other oxidative stress induced TFs are activated in the first phase with FOXO1 (NF-κB, NFAT1), or the second phase with p53 (NRF2, JUN) but not both following H2O2stress. The two TF phases result in large differences in gene expression patterns. Finally, we provide evidence that 2-Cys peroxiredoxins control the timing of the TF phases in response to H2O2.
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| Overall design |
MCF7 cells (50,000 cells/well) were plated on plastic 6 well plates and allowed to attach for 2 days. There were nine different treatment groups: MCF7 + PBS Control, MCF7 + 50 µM H2O2, MCF7 + 75 µM H2O2, MCF7 + 20 µM J14 + PBS Control, MCF7 + 20 µM J14 + 50 µM H2O2, MCF7 + 20 µM J14 + 75 µM H2O2, MCF7 SFRX-OE + PBS Control, MCF7 SFRX-OE + 50 µM H2O2, MCF7 SFRX-OE + 75 µM H2O2. SFRX-OE indicate the cell line harbored a construct that expressed the human Sulfiredoxin gene from the PGK promoter. J14 is a Sulfiredoxin inhibitor. Five hours after treatment, RNA was isolated using RNeasy Mini Kit (Cat. No. 74104) from Qiagen. The isolated RNA was then sent to Novogene for sequencing.
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| Contributor(s) |
Paek A, Jose E, March-Steinman W |
| Citation(s) |
38653977 |
| |
| Submission date |
Mar 16, 2023 |
| Last update date |
May 17, 2024 |
| Contact name |
Andrew Paek |
| E-mail(s) |
apaek@arizona.edu
|
| Phone |
2102942343
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| Organization name |
University of Arizona
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| Department |
Molecular and Cellular Biology
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| Street address |
Life Sciences South, 1007 E. Lowell Stre, PO Box 210106
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| City |
Tucson |
| State/province |
Arizona |
| ZIP/Postal code |
85721 |
| Country |
USA |
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| Platforms (1) |
| GPL24676 |
Illumina NovaSeq 6000 (Homo sapiens) |
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| Samples (27)
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| This SubSeries is part of SuperSeries: |
| GSE227556 |
Temporal Coordination of the Transcription Factor Response to H2O2stress |
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| Relations |
| BioProject |
PRJNA945497 |
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