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α-ketoglutarate coordinates carbon and nitrogen utilization via enzyme I inhibition

Nature Chemical Biology volume 7, pages 894–901 (2011)Cite this article

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Abstract

Microbes survive in a variety of nutrient environments by modulating their intracellular metabolism. Balanced growth requires coordinated uptake of carbon and nitrogen, the primary substrates for biomass production. Yet the mechanisms that balance carbon and nitrogen uptake are poorly understood. We find in Escherichia coli that a sudden increase in nitrogen availability results in an almost immediate increase in glucose uptake. The concentrations of glycolytic intermediates and known regulators, however, remain homeostatic. Instead, we find that α-ketoglutarate, which accumulates in nitrogen limitation, directly blocks glucose uptake by inhibiting enzyme I, the first step of the sugar–phosphoenolpyruvate phosphotransferase system (PTS). This inhibition enables rapid modulation of glycolytic flux without marked changes in the concentrations of glycolytic intermediates by simultaneously altering import of glucose and consumption of the terminal glycolytic intermediate phosphoenolpyruvate. Quantitative modeling shows that this previously unidentified regulatory connection is, in principle, sufficient to coordinate carbon and nitrogen utilization.

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Figure 1: Glycolytic intermediates are homeostatic during nitrogen upshift.
Figure 2: Glucose uptake increases rapidly upon nitrogen upshift, independent of glutamine concentration.
Figure 3: α-ketoglutarate inhibits enzyme I in vitro.
Figure 4: Simulated pool size changes in response to varying nitrogen availability.

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Acknowledgements

The authors thank J. Yuan, whose experiments laid the groundwork for the current study; D. Yan for providing the GOGAT strain; and G. Gosset for providing the W3110 and VH33 strains. This research was funded by US National Science Foundation CAREER award MCB-0643859, joint Department of Energy–Air Force Office of Scientific Research Award DOE DE-SC0002077–AFOSR FA9550-09-1-0580, and the US National Institutes of Health Center for Quantitative Biology Award P50 GM071508.

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Authors and Affiliations

  1. Lewis-Sigler Institute for Integrative Genomics, Princeton University, Princeton, New Jersey, USA

    Christopher D Doucette, David J Schwab, Ned S Wingreen & Joshua D Rabinowitz

  2. Department of Molecular Biology, Princeton University, Princeton, New Jersey, USA

    Christopher D Doucette, David J Schwab & Ned S Wingreen

  3. Department of Chemistry, Princeton University, Princeton, New Jersey, USA

    Joshua D Rabinowitz

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C.D.D. designed and performed experiments, designed the model and wrote the paper; D.J.S. designed the model and performed the model analysis; N.S.W. designed the model; J.D.R. designed experiments, designed the model and wrote the paper.

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Correspondence to Joshua D Rabinowitz.

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Doucette, C., Schwab, D., Wingreen, N. et al. α-ketoglutarate coordinates carbon and nitrogen utilization via enzyme I inhibition. Nat Chem Biol 7, 894–901 (2011). https://doi.org/10.1038/nchembio.685

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