PGC-1β and ERRα Promote Glutamine Metabolism and Colorectal Cancer Survival via Transcriptional Upregulation of PCK2
Abstract
:Simple Summary
Abstract
1. Introduction
2. Materials and Methods
2.1. Cell Culture
2.2. Lentiviral Transduction
2.3. Epitope-Tagging of Endogenous PGC-1β
2.4. Immunoprecipitation of PGC-1β
2.5. Inducible PGC-1β Vector and Cell Line Generation
2.6. siRNA Transfections
2.7. RNA Sequencing and Analysis
2.8. Western Blot Analyses
2.9. L-Glutamine Utilization Assays
2.10. Intracellular Metabolite Analysis
2.11. PCK2-ALFA Plasmid for Stable Expression
2.12. Confocal Microscopy
2.13. Statistical Analysis
3. Results
3.1. Endogenous PGC-1β Interacts with ERRα, Promotes ERRα Expression, and Anchorage-Independent Growth
3.2. PGC-1β Requires Its LRELL Motif at Amino Acids 343–347 to Interact with ERRα
3.3. PGC-1β and ERRα Promote PCK2 Expression
3.4. PCK2 Promotes Anchorage-Independent Growth and Glutamine Utilization
3.5. ERRα Depletion Causes Loss of Anchorage-Independent Growth and Glutamine Utilization That Is Not Rescued by Over-Expression of PCK2
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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Frodyma, D.E.; Troia, T.C.; Rao, C.; Svoboda, R.A.; Berg, J.A.; Shinde, D.D.; Thomas, V.C.; Lewis, R.E.; Fisher, K.W. PGC-1β and ERRα Promote Glutamine Metabolism and Colorectal Cancer Survival via Transcriptional Upregulation of PCK2. Cancers 2022, 14, 4879. https://doi.org/10.3390/cancers14194879
Frodyma DE, Troia TC, Rao C, Svoboda RA, Berg JA, Shinde DD, Thomas VC, Lewis RE, Fisher KW. PGC-1β and ERRα Promote Glutamine Metabolism and Colorectal Cancer Survival via Transcriptional Upregulation of PCK2. Cancers. 2022; 14(19):4879. https://doi.org/10.3390/cancers14194879
Chicago/Turabian StyleFrodyma, Danielle E., Thomas C. Troia, Chaitra Rao, Robert A. Svoboda, Jordan A. Berg, Dhananjay D. Shinde, Vinai C. Thomas, Robert E. Lewis, and Kurt W. Fisher. 2022. "PGC-1β and ERRα Promote Glutamine Metabolism and Colorectal Cancer Survival via Transcriptional Upregulation of PCK2" Cancers 14, no. 19: 4879. https://doi.org/10.3390/cancers14194879