PKM2 functions as a histidine kinase to phosphorylate PGAM1 and increase glycolysis shunts in cancer.

Phosphoglycerate mutase 1 (PGAM1) is a key node enzyme that diverts the metabolic reactions from glycolysis into its shunts to support macromolecule biosynthesis for rapid and sustainable cell proliferation. It is prevalent that PGAM1 activity is upregulated in various tumors; however, the underlying mechanism remains unclear. Here, we unveil that pyruvate kinase M2 (PKM2) moonlights as a histidine kinase in a phosphoenolpyruvate (PEP)-dependent manner to catalyze PGAM1 H11 phosphorylation, that is essential for PGAM1 activity. Moreover, monomeric and dimeric but not tetrameric PKM2 are efficient to phosphorylate and activate PGAM1. In response to epidermal growth factor signaling, Src-catalyzed PGAM1 Y119 phosphorylation is a prerequisite for PKM2 binding and the subsequent PGAM1 H11 phosphorylation, which constitutes a discrepancy between tumor and normal cells. A PGAM1-derived pY119-containing cell-permeable peptide or Y119 mutation disrupts the interaction of PGAM1 with PKM2 and PGAM1 H11 phosphorylation, dampening the glycolysis shunts and tumor growth. Together, these results identify a function of PKM2 as a histidine kinase, and illustrate the importance of enzyme crosstalk as a regulatory mode during metabolic reprogramming and tumorigenesis. Synopsis: The mechanistic contribution of phosphoglycerate mutase 1 (PGAM1), a metabolic enzyme active in glycolysis to tumorigenesis remains unclear. Here, PGAM1 activity is shown to be controlled by pyruvate kinase M2 (PKM2), promoting glycolytic shunts and macromolecule biosynthesis during metabolic reprogramming in cancer cells. PKM2 moonlights as a histidine kinase and phosphorylates PGAM1 at H11 by transferring phosphate from phosphoenolpyruvate. De-tetramerization is favorable for PKM2 functioning as a histidine kinase, but disadvantageous for its glycolytic activity. PGAM1 Y119 phosphorylation induced by EGF-Src signaling is required for PKM2 recognition and binding. A PGAM1-derived pY119-containing cell-permeable peptide blocks PGAM1-PKM2 interaction, thereby slowing down tumor growth. Non-canonical PKM2 function controls PGAM1 and macromolecule biosynthesis during metabolic reprogramming and tumor growth. [ABSTRACT FROM AUTHOR]