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Title: NADPH production by the oxidative pentose-phosphate pathway supports folate metabolism

Abstract

NADPH donates high-energy electrons for antioxidant defence and reductive biosynthesis. Cytosolic NADP is recycled to NADPH by the oxidative pentose-phosphate pathway (oxPPP), malic enzyme 1 (ME1) and isocitrate dehydrogenase 1 (IDH1). Here we show that any one of these routes can support cell growth, but the oxPPP is uniquely required to maintain a normal NADPH/NADP ratio, mammalian dihydrofolate reductase (DHFR) activity and folate metabolism. These findings are based on CRISPR deletions of glucose-6-phosphate dehydrogenase (G6PD, the committed oxPPP enzyme), ME1, IDH1 and combinations thereof in HCT116 colon cancer cells. Loss of G6PD results in high NADP, which induces compensatory increases in ME1 and IDH1 flux. But the high NADP inhibits DHFR, resulting in impaired folate-mediated biosynthesis, which is reversed by recombinant expression of Escherichia coli DHFR. Conclusively, across different cancer cell lines, G6PD deletion produced consistent changes in folate-related metabolites, suggesting a general requirement for the oxPPP to support folate metabolism.

Authors:
ORCiD logo [1];  [1];  [2];  [1]; ORCiD logo [2];  [2]; ORCiD logo [1]
  1. Princeton Univ., Princeton, NJ (United States)
  2. Univ. of Pennsylvania, Philadelphia, PA (United States)
Publication Date:
Research Org.:
Center for Advanced Bioenergy and Bioproducts Innovation (CABBI), Urbana, IL (United States)
Sponsoring Org.:
USDOE Office of Science (SC), Biological and Environmental Research (BER) (SC-23)
OSTI Identifier:
1501460
Grant/Contract Number:  
SC0018420
Resource Type:
Accepted Manuscript
Journal Name:
Nature Metabolism
Additional Journal Information:
Journal Volume: 1; Journal Issue: 3; Journal ID: ISSN 2522-5812
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES

Citation Formats

Chen, Li, Zhang, Zhaoyue, Hoshino, Atsushi, Zheng, Henry D., Morley, Michael, Arany, Zoltan, and Rabinowitz, Joshua D. NADPH production by the oxidative pentose-phosphate pathway supports folate metabolism. United States: N. p., 2019. Web. doi:10.1038/s42255-019-0043-x.
Chen, Li, Zhang, Zhaoyue, Hoshino, Atsushi, Zheng, Henry D., Morley, Michael, Arany, Zoltan, & Rabinowitz, Joshua D. NADPH production by the oxidative pentose-phosphate pathway supports folate metabolism. United States. doi:10.1038/s42255-019-0043-x.
Chen, Li, Zhang, Zhaoyue, Hoshino, Atsushi, Zheng, Henry D., Morley, Michael, Arany, Zoltan, and Rabinowitz, Joshua D. Mon . "NADPH production by the oxidative pentose-phosphate pathway supports folate metabolism". United States. doi:10.1038/s42255-019-0043-x. https://www.osti.gov/servlets/purl/1501460.
@article{osti_1501460,
title = {NADPH production by the oxidative pentose-phosphate pathway supports folate metabolism},
author = {Chen, Li and Zhang, Zhaoyue and Hoshino, Atsushi and Zheng, Henry D. and Morley, Michael and Arany, Zoltan and Rabinowitz, Joshua D.},
abstractNote = {NADPH donates high-energy electrons for antioxidant defence and reductive biosynthesis. Cytosolic NADP is recycled to NADPH by the oxidative pentose-phosphate pathway (oxPPP), malic enzyme 1 (ME1) and isocitrate dehydrogenase 1 (IDH1). Here we show that any one of these routes can support cell growth, but the oxPPP is uniquely required to maintain a normal NADPH/NADP ratio, mammalian dihydrofolate reductase (DHFR) activity and folate metabolism. These findings are based on CRISPR deletions of glucose-6-phosphate dehydrogenase (G6PD, the committed oxPPP enzyme), ME1, IDH1 and combinations thereof in HCT116 colon cancer cells. Loss of G6PD results in high NADP, which induces compensatory increases in ME1 and IDH1 flux. But the high NADP inhibits DHFR, resulting in impaired folate-mediated biosynthesis, which is reversed by recombinant expression of Escherichia coli DHFR. Conclusively, across different cancer cell lines, G6PD deletion produced consistent changes in folate-related metabolites, suggesting a general requirement for the oxPPP to support folate metabolism.},
doi = {10.1038/s42255-019-0043-x},
journal = {Nature Metabolism},
number = 3,
volume = 1,
place = {United States},
year = {2019},
month = {3}
}

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Works referenced in this record:

Idh1 protects murine hepatocytes from endotoxin-induced oxidative stress by regulating the intracellular NADP+/NADPH ratio
journal, April 2015

  • Itsumi, M.; Inoue, S.; Elia, A. J.
  • Cell Death & Differentiation, Vol. 22, Issue 11
  • DOI: 10.1038/cdd.2015.38

Compartmentalization of Mammalian Folate-Mediated One-Carbon Metabolism
journal, July 2010


Evaluation of 13C isotopic tracers for metabolic flux analysis in mammalian cells
journal, November 2009

  • Metallo, Christian M.; Walther, Jason L.; Stephanopoulos, Gregory
  • Journal of Biotechnology, Vol. 144, Issue 3
  • DOI: 10.1016/j.jbiotec.2009.07.010

One-Carbon Metabolism in Health and Disease
journal, January 2017


A domino effect in antifolate drug action in Escherichia coli
journal, August 2008

  • Kwon, Yun Kyung; Lu, Wenyun; Melamud, Eugene
  • Nature Chemical Biology, Vol. 4, Issue 10
  • DOI: 10.1038/nchembio.108

Reductive carboxylation supports redox homeostasis during anchorage-independent growth
journal, April 2016

  • Jiang, Lei; Shestov, Alexander A.; Swain, Pamela
  • Nature, Vol. 532, Issue 7598
  • DOI: 10.1038/nature17393

Serine, glycine and one-carbon units: cancer metabolism in full circle
journal, July 2013

  • Locasale, Jason W.
  • Nature Reviews Cancer, Vol. 13, Issue 8
  • DOI: 10.1038/nrc3557

Oxidative stress inhibits distant metastasis by human melanoma cells
journal, October 2015

  • Piskounova, Elena; Agathocleous, Michalis; Murphy, Malea M.
  • Nature, Vol. 527, Issue 7577
  • DOI: 10.1038/nature15726

Aerobic Glycolysis: Meeting the Metabolic Requirements of Cell Proliferation
journal, November 2011


An embryoprotective role for glucose-6-phosphate dehydrogenase in developmental oxidative stress and chemical teratogenesis
journal, January 2000

  • Nicol, Christopher J.; Zielenski, Julian; Tsui, Lap-Chee
  • The FASEB Journal, Vol. 14, Issue 1
  • DOI: 10.1096/fasebj.14.1.111

Malic enzyme tracers reveal hypoxia-induced switch in adipocyte NADPH pathway usage
journal, March 2016

  • Liu, Ling; Shah, Supriya; Fan, Jing
  • Nature Chemical Biology, Vol. 12, Issue 5
  • DOI: 10.1038/nchembio.2047

Functional significance of evolving protein sequence in dihydrofolate reductase from bacteria to humans
journal, June 2013

  • Liu, C. T.; Hanoian, P.; French, J. B.
  • Proceedings of the National Academy of Sciences, Vol. 110, Issue 25
  • DOI: 10.1073/pnas.1307130110

Reversal of Cytosolic One-Carbon Flux Compensates for Loss of the Mitochondrial Folate Pathway
journal, June 2016


Improved vectors and genome-wide libraries for CRISPR screening
journal, July 2014

  • Sanjana, Neville E.; Shalem, Ophir; Zhang, Feng
  • Nature Methods, Vol. 11, Issue 8
  • DOI: 10.1038/nmeth.3047

Chemical Basis for Deuterium Labeling of Fat and NADPH
journal, October 2017

  • Zhang, Zhaoyue; Chen, Li; Liu, Ling
  • Journal of the American Chemical Society, Vol. 139, Issue 41
  • DOI: 10.1021/jacs.7b08012

Cytosolic NADP + -dependent Isocitrate Dehydrogenase Plays a Key Role in Lipid Metabolism
journal, July 2004

  • Koh, Ho-Jin; Lee, Su-Min; Son, Byung-Gap
  • Journal of Biological Chemistry, Vol. 279, Issue 38
  • DOI: 10.1074/jbc.M402260200

Serine Catabolism Regulates Mitochondrial Redox Control during Hypoxia
journal, September 2014


Genome engineering using the CRISPR-Cas9 system
journal, October 2013


Glucose feeds the TCA cycle via circulating lactate
journal, October 2017

  • Hui, Sheng; Ghergurovich, Jonathan M.; Morscher, Raphael J.
  • Nature, Vol. 551, Issue 7678
  • DOI: 10.1038/nature24057

Mitochondrial One-Carbon Pathway Supports Cytosolic Folate Integrity in Cancer Cells
journal, November 2018


Glucose-6-phosphate dehydrogenase deficiency
journal, January 2008


Reductive glutamine metabolism by IDH1 mediates lipogenesis under hypoxia
journal, November 2011

  • Metallo, Christian M.; Gameiro, Paulo A.; Bell, Eric L.
  • Nature, Vol. 481, Issue 7381
  • DOI: 10.1038/nature10602

Human mutations in methylenetetrahydrofolate dehydrogenase 1 impair nuclear de novo thymidylate biosynthesis
journal, December 2014

  • Field, Martha S.; Kamynina, Elena; Watkins, David
  • Proceedings of the National Academy of Sciences, Vol. 112, Issue 2
  • DOI: 10.1073/pnas.1414555112

Reciprocal regulation of p53 and malic enzymes modulates metabolism and senescence
journal, January 2013

  • Jiang, Peng; Du, Wenjing; Mancuso, Anthony
  • Nature, Vol. 493, Issue 7434
  • DOI: 10.1038/nature11776

Quantitative flux analysis reveals folate-dependent NADPH production
journal, May 2014

  • Fan, Jing; Ye, Jiangbin; Kamphorst, Jurre J.
  • Nature, Vol. 510, Issue 7504
  • DOI: 10.1038/nature13236

Hypoxia promotes isocitrate dehydrogenase-dependent carboxylation of  -ketoglutarate to citrate to support cell growth and viability
journal, November 2011

  • Wise, D. R.; Ward, P. S.; Shay, J. E. S.
  • Proceedings of the National Academy of Sciences, Vol. 108, Issue 49
  • DOI: 10.1073/pnas.1117773108

Metabolite Spectral Accuracy on Orbitraps
journal, May 2017


Spatial distribution of G6PD deficiency variants across malaria-endemic regions
journal, November 2013

  • Howes, Rosalind E.; Dewi, Mewahyu; Piel, Frédéric B.
  • Malaria Journal, Vol. 12, Issue 1
  • DOI: 10.1186/1475-2875-12-418

A roadmap for interpreting 13 C metabolite labeling patterns from cells
journal, August 2015


p53 regulates biosynthesis through direct inactivation of glucose-6-phosphate dehydrogenase
journal, February 2011

  • Jiang, Peng; Du, Wenjing; Wang, Xingwu
  • Nature Cell Biology, Vol. 13, Issue 3
  • DOI: 10.1038/ncb2172

Metabolomic Analysis and Visualization Engine for LC−MS Data
journal, December 2010

  • Melamud, Eugene; Vastag, Livia; Rabinowitz, Joshua D.
  • Analytical Chemistry, Vol. 82, Issue 23
  • DOI: 10.1021/ac1021166

An LC-MS chemical derivatization method for the measurement of five different one-carbon states of cellular tetrahydrofolate
journal, August 2017

  • Chen, Li; Ducker, Gregory S.; Lu, Wenyun
  • Analytical and Bioanalytical Chemistry, Vol. 409, Issue 25
  • DOI: 10.1007/s00216-017-0514-4

Tracing Compartmentalized NADPH Metabolism in the Cytosol and Mitochondria of Mammalian Cells
journal, July 2014


Serine and one-carbon metabolism in cancer
journal, September 2016

  • Yang, Ming; Vousden, Karen H.
  • Nature Reviews Cancer, Vol. 16, Issue 10
  • DOI: 10.1038/nrc.2016.81

Metabolism of Citrate and Other Carboxylic Acids in Erythrocytes As a Function of Oxygen Saturation and Refrigerated Storage
journal, October 2017