Abstract
Lower glycolysis involves a series of reversible reactions, which interconvert intermediates that also feed anabolic pathways. 3-phosphoglycerate (3-PG) is an abundant lower glycolytic intermediate that feeds serine biosynthesis via the enzyme phosphoglycerate dehydrogenase, which is genomically amplified in several cancers. Phosphoglycerate mutase 1 (PGAM1) catalyzes the isomerization of 3-PG into the downstream glycolytic intermediate 2-phosphoglycerate (2-PG). PGAM1 needs to be histidine phosphorylated to become catalytically active. We show that the primary PGAM1 histidine phosphate donor is 2,3-bisphosphoglycerate (2,3-BPG), which is made from the glycolytic intermediate 1,3-bisphosphoglycerate (1,3-BPG) by bisphosphoglycerate mutase (BPGM). When BPGM is knocked out, 1,3-BPG can directly phosphorylate PGAM1. In this case, PGAM1 phosphorylation and activity are decreased, but nevertheless sufficient to maintain normal glycolytic flux and cellular growth rate. 3-PG, however, accumulates, leading to increased serine synthesis. Thus, one biological function of BPGM is controlling glycolytic intermediate levels and thereby serine biosynthetic flux.
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Acknowledgements
We thank current and former members of the Muir and Rabinowitz laboratories for helpful discussions during the preparation of this manuscript and V. Suri of Raze Therapeutics for technical help. This work was funded by the US National Institutes of Health for T.W.M. (5R01GM095880) and J.D.R. (R01 CA163591 and P30DK019525), the US Department of Energy for J.D.R. (DE-SC0012461), Stand Up to Cancer for J.D.R. (SU2C-AACR-DT0509), and the Brewster Foundation and Breast Cancer Research Foundation for Y.K. R.C.O. and J.-M.K. were supported by postdoctoral fellowships from the National Institutes of Health Research Service Award (1F32CA167901) and Damon Runyon Cancer Research Foundation (DRG-2005-09), respectively.
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R.C.O. designed experiments, prepared materials, analyzed data, interpreted results, generated HEK 293T and HCT116 knockout cell lines, and conducted experiments for metabolomic data analysis, western blotting, cell growth measurements, and in vitro phosphorylation analysis. X.S. designed experiments, prepared materials, analyzed data, interpreted results, designed primers and performed all DNA sequencing analysis for BPGM knockout experiments, conducted all metabolomic experiments and data analysis, prepared 1,3-BPG, and derived and conducted all serine flux calculations. M.H. designed experiments, analyzed data, interpreted results, performed cell assays for metabolomic data analysis, performed western blots for BPGM rescue and PGAM1 knockdown experiments, and performed cell growth measurements of knockout cells with serine/glycine or oxygen depletion. J.-M.K. designed experiments, analyzed data, interpreted results, and performed western blot analysis of BPGM protein levels. M.E. generated MDA-MB-231 knockout cells, prepared PGAM1 knockdown cells, and designed, performed, and analyzed mouse xenograft experiments. Y.D. designed and conducted RNA-seq analysis of wild-type and knockout HEK 293T cells. B.W. designed and conducted 32P-labeling experiments, analyzed data, and interpreted results of metabolomic experiments. E.G. performed phosphorylated PGAM1 western blot analysis from multiple cell lines. D.H.P. performed LC–MS-based proteomic analysis of phosphorylated PGAM1, analyzed data, and interpreted results. T.W.M. designed experiments, analyzed data, and interpreted results. J.D.R. designed experiments, analyzed data, and interpreted results. R.C.O., X.S., M.H., T.W.M., and J.D.R. wrote the manuscript with contributions from all authors.
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Oslund, R., Su, X., Haugbro, M. et al. Bisphosphoglycerate mutase controls serine pathway flux via 3-phosphoglycerate. Nat Chem Biol 13, 1081–1087 (2017). https://doi.org/10.1038/nchembio.2453
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DOI: https://doi.org/10.1038/nchembio.2453
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