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ULK1-dependent phosphorylation of PKM2 antagonizes O-GlcNAcylation and regulates the Warburg effect in breast cancer

Abstract

Pyruvate kinase M2 (PKM2) is a central metabolic enzyme driving the Warburg effect in tumor growth. Previous investigations have demonstrated that PKM2 is subject to O-linked β-N-acetylglucosamine (O-GlcNAc) modification, which is a nutrient-sensitive post-translational modification. Here we found that unc-51 like autophagy activating kinase 1 (ULK1), a glucose-sensitive kinase, interacts with PKM2 and phosphorylates PKM2 at Ser333. Ser333 phosphorylation antagonizes PKM2 O-GlcNAcylation, promotes its tetramer formation and enzymatic activity, and decreases its nuclear localization. As PKM2 is known to have a nuclear role in regulating c-Myc, we also show that PKM2-S333 phosphorylation inhibits c-Myc expression. By downregulating glucose consumption and lactate production, PKM2 pS333 attenuates the Warburg effect. Through mouse xenograft assays, we demonstrate that the phospho-deficient PKM2-S333A mutant promotes tumor growth in vivo. In conclusion, we identified a ULK1-PKM2-c-Myc axis in inhibiting breast cancer, and a glucose-sensitive phosphorylation of PKM2 in modulating the Warburg effect.

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Fig. 1: ULK1 interacts with PKM2.
Fig. 2: ULK1 phosphorylates PKM2 at pS333.
Fig. 3: PKM2 pS333 antagonizes O-GlcNAcylation.
Fig. 4: PKM2 pS333 reduces pS37 levels and decreases nuclear localization.
Fig. 5: Abrogation of PKM2 pS333 increases the Warburg effect.
Fig. 6: Abrogation of PKM2 pS333 decreases tetramer formation enzymatic activity.
Fig. 7: Abrogation of PKM2 pS333 enhances breast cancer progression.
Fig. 8: Model depicting the role of PKM2 S333 phosphorylation.

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Data availability

The mass spectrometry proteomics data have been deposited to the ProteomeXchange Consortium (http://proteomecentral.proteomexchange.org) via the iProX partner repository [25, 26] with the dataset identifier PXD044583.

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Acknowledgements

We thank Dr. Hong-Jie Zhang (Core Facility for Protein Research, Institute of Biophysics, Chinese Academy of Sciences) for the technique support using radioactivity detection. JL is supported by the National Natural Science Foundation of China (NSFC) fund (32271285 and 31872720) and R & D Program of Beijing Municipal Education Commission (KZ202210028043). XX is supported by NSFC fund 32090031, the Shenzhen Science and Technology Innovation Commission projects JCYJ201805073000163.

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Zibin Zhou: investigation; Xiyuan Zheng: investigation; Jianxin Zhao: investigation; Aiyun Yuan: investigation; Zhuan Lv: investigation; Guangcan Shao: investigation; Bin Peng: investigation; Meng-Qiu Dong: Resources, Supervision; Quan Xu: conceptualization, data curation, supervision, project administration; Xingzhi Xu: conceptualization, resources, supervision, project administration; Jing Li: conceptualization, supervision, project administration, funding acquisition.

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Correspondence to Quan Xu, Xingzhi Xu or Jing Li.

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Zhou, Z., Zheng, X., Zhao, J. et al. ULK1-dependent phosphorylation of PKM2 antagonizes O-GlcNAcylation and regulates the Warburg effect in breast cancer. Oncogene 43, 1769–1778 (2024). https://doi.org/10.1038/s41388-024-03035-y

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