Abstract
The cystine/glutamate antiporter SLC7A11 (commonly known as xCT) functions to import cystine for glutathione biosynthesis, thereby protecting cells from oxidative stress and ferroptosis, a regulated form of non-apoptotic cell death driven by the accumulation of lipid-based reactive oxygen species (ROS). p14ARF, a well-established tumor suppressor, promotes ferroptosis by inhibiting NRF2-mediated SLC7A11 transcription. Here, we demonstrate the crucial role of Cullin 2 RING E3 ligase (CRL2)-KLHDC3 E3 ubiquitin ligase complex in regulating p14ARF protein stability. KLHDC3 acts as a CRL2 adaptor that specifically recognizes a C-terminal degron in p14ARF and triggers p14ARF for ubiquitin–proteasomal degradation. This regulation mode is absent in the murine p14ARF homolog, p19arf which lacks the C-terminal degron. We also show that KLHDC3 suppresses ferroptosis in vitro and supports tumor growth in vivo by relieving p14ARF-mediated suppression of SLC7A11 transcription. Overall, these findings reveal that the protein stability and pro-ferroptotic function of p14ARF are controlled by a CRL2 E3 ubiquitin ligase complex, and suggest that suppression of the p14ARF-NRF2-SLC7A11 regulatory pathway by KLHDC3 overexpression likely contributes to cancer progression.
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Data availability
Detailed information on the reagents, sequences of primers and sgRNAs s can be found in Supplementary Tables 1–3. For original data, please contact [email protected].
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Acknowledgements
We thank Dr. Lin Huang for excellent technical assistance in mass spectrometry analysis
Funding
This work was in part supported by the National Natural Science Foundation of China (nos. 81872260, 82172938 to PZ; nos. 91957125, 81972396, to CW; nos. 91954106, 81872109 to KG).
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PZZ, CJW, and GW conceived the study. PZZ, KG, QS, XYZ, LZ, YJC, DYJ, ZL and HRS performed the experiments and data analyses. PZZ, KG, YL, and LZ analyzed and interpreted the data. CJW and PZZ wrote the paper.
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Our studies did not include human participants or human tissue. The animal studies were approved by ethics Review Committee for Animal Experimentation of Fudan University.
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Zhang, P., Gao, K., Zhang, L. et al. CRL2-KLHDC3 E3 ubiquitin ligase complex suppresses ferroptosis through promoting p14ARF degradation. Cell Death Differ 29, 758–771 (2022). https://doi.org/10.1038/s41418-021-00890-0
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DOI: https://doi.org/10.1038/s41418-021-00890-0
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