Ochratoxin A Sequentially Activates Autophagy and the Ubiquitin-Proteasome System
Abstract
:1. Introduction
2. Results
2.1. OTA Promotes Autophagic Activity in HK-2 Cells
2.2. OTA-Induced Autophagic Activity Has a Pro-Death Effect in MEF Cells
2.3. OTA Promotes Proteasomal Degradation As a Late Response in HK-2 and MEF Cells
2.4. OTA-Induced UPS Activity is Not Related to the Levels of the Proteasome Subunits
2.5. OTA Directly Increases the Proteolytic Activities of the 26S But Not the 20S Proteasome
2.6. OTA Stimulates Degradation of the Exogenous UPS Substrate, Ubiquitinated-Green Fluorescence Protein (Ub-GFP)
2.7. OTA Decreases the Half-Lives of the Natural Substrate Proteins of UPS; MCL1, KEAP1, and IKBα
2.8. OTA Facilitates Degradation of Phosphatases Involved in PI3K/AKT and MAPK/ERK1-2 Pathways Through Activation of Autophagy and UPS
3. Discussion
4. Conclusions
5. Materials and Methods
5.1. Cell Culture
5.2. Stable Cell Line
5.3. Chemicals and Treatments
5.4. Cell Viability Assay
5.5. Protein Extraction and Western Blot Analysis
5.6. Detection of Autophagy and Acidic Vesicles upon OTA Treatment in HK-2 Cell Line
5.7. Measurement of Proteasome Activity
5.8. Detection of the Expression Levels of the Components of 26S Proteasome by RT-qPCR
5.9. Transient Transfection of the HeLa Cells with Ub-GFP Plasmids, Visualization under Fluorescence Microscope, and Flow Cytometric Analysis
5.10. Statistical Analysis
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Akpinar, H.A.; Kahraman, H.; Yaman, I. Ochratoxin A Sequentially Activates Autophagy and the Ubiquitin-Proteasome System. Toxins 2019, 11, 615. https://doi.org/10.3390/toxins11110615
Akpinar HA, Kahraman H, Yaman I. Ochratoxin A Sequentially Activates Autophagy and the Ubiquitin-Proteasome System. Toxins. 2019; 11(11):615. https://doi.org/10.3390/toxins11110615
Chicago/Turabian StyleAkpinar, Hafize Aysin, Hilal Kahraman, and Ibrahim Yaman. 2019. "Ochratoxin A Sequentially Activates Autophagy and the Ubiquitin-Proteasome System" Toxins 11, no. 11: 615. https://doi.org/10.3390/toxins11110615