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:
-
[1];
[2];
[1]
- Princeton Univ., Princeton, NJ (United States)
- 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)
- 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. https://doi.org/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. https://doi.org/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 = {Mon Mar 11 00:00:00 EDT 2019},
month = {Mon Mar 11 00:00:00 EDT 2019}
}
Free Publicly Available Full Text
Publisher's Version of Record
Other availability
Search WorldCat to find libraries that may hold this journal
Save to My Library
You must Sign In or Create an Account in order to save documents to your library.
Works referenced in this record:
Compartmentalization of Mammalian Folate-Mediated One-Carbon Metabolism
journal, July 2010
- Tibbetts, Anne S.; Appling, Dean R.
- Annual Review of Nutrition, Vol. 30, Issue 1
The role of metabolic reprogramming and de novo amino acid synthesis in collagen protein production by myofibroblasts: implications for organ fibrosis and cancer
journal, May 2021
- Hamanaka, Robert B.; Mutlu, Gökhan M.
- Amino Acids
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
Reversal of Cytosolic One-Carbon Flux Compensates for Loss of the Mitochondrial Folate Pathway
journal, June 2016
- Ducker, Gregory S.; Chen, Li; Morscher, Raphael J.
- Cell Metabolism, Vol. 23, Issue 6
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
Serine Catabolism Regulates Mitochondrial Redox Control during Hypoxia
journal, September 2014
- Ye, J.; Fan, J.; Venneti, S.
- Cancer Discovery, Vol. 4, Issue 12
Glucose feeds the TCA cycle via circulating lactate
journal, October 2017
- Hui, Sheng; Ghergurovich, Jonathan M.; Morscher, Raphael J.
- Nature, Vol. 551, Issue 7678
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
Cytosolic Malic Enzyme 1 (ME1) Mediates High Fat Diet-Induced Adiposity, Endocrine Profile, and Gastrointestinal Tract Proliferation-Associated Biomarkers in Male Mice
journal, October 2012
- Al-Dwairi, Ahmed; Pabona, John Mark P.; Simmen, Rosalia C. M.
- PLoS ONE, Vol. 7, Issue 10
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
Glucose-6-Phosphate Dehydrogenase Deficiency
journal, October 1981
- Bienzle, U.
- Clinics in Haematology, Vol. 10, Issue 3
A roadmap for interpreting 13 C metabolite labeling patterns from cells
journal, August 2015
- Buescher, Joerg M.; Antoniewicz, Maciek R.; Boros, Laszlo G.
- Current Opinion in Biotechnology, Vol. 34
Reversal of Cytosolic One-Carbon Flux Compensates for Loss of the Mitochondrial Folate Pathway
text, January 2016
- Ducker, Gregory S.; Chen, Li; Morscher, Raphael J.
- Cell Press (Elsevier)
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
The Labeling of Pentose Phosphate from Glucose- 14 C and Estimation of the Rates of Transaldolase, Transketolase, the Contribution of the Pentose Cycle, and Ribose Phosphate Synthesis *
journal, July 1967
- Katz, Joseph; Rognstad, Robert
- Biochemistry, Vol. 6, Issue 7
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
Glucose feeds the TCA cycle via circulating lactate
text, January 2017
- Hui, Sheng; Ghergurovich, Jonathan M.; Morscher, Raphael J.
- Nature Publishing Group
Glucose-6-phosphate dehydrogenase deficiency.
journal, December 1994
- Mehta, A. B.
- Postgraduate Medical Journal, Vol. 70, Issue 830
Metabolism of Citrate and Other Carboxylic Acids in Erythrocytes As a Function of Oxygen Saturation and Refrigerated Storage
journal, October 2017
- Nemkov, Travis; Sun, Kaiqi; Reisz, Julie A.
- Frontiers in Medicine, Vol. 4
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
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
One-Carbon Metabolism in Health and Disease
journal, January 2017
- Ducker, Gregory S.; Rabinowitz, Joshua D.
- Cell Metabolism, Vol. 25, Issue 1
Reductive carboxylation supports redox homeostasis during anchorage-independent growth
journal, April 2016
- Jiang, Lei; Shestov, Alexander A.; Swain, Pamela
- Nature, Vol. 532, Issue 7598
Serine, glycine and one-carbon units: cancer metabolism in full circle
journal, July 2013
- Locasale, Jason W.
- Nature Reviews Cancer, Vol. 13, Issue 8
Oxidative stress inhibits distant metastasis by human melanoma cells
journal, October 2015
- Piskounova, Elena; Agathocleous, Michalis; Murphy, Malea M.
- Nature, Vol. 527, Issue 7577
Aerobic Glycolysis: Meeting the Metabolic Requirements of Cell Proliferation
journal, November 2011
- Lunt, Sophia Y.; Vander Heiden, Matthew G.
- Annual Review of Cell and Developmental Biology, Vol. 27, Issue 1
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
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
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
Reversal of Cytosolic One-Carbon Flux Compensates for Loss of the Mitochondrial Folate Pathway
journal, October 2016
- Ducker, Gregory S.; Chen, Li; Morscher, Raphael J.
- Cell Metabolism, Vol. 24, Issue 4
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
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
trans-Fatty acids promote p53-dependent apoptosis triggered by cisplatin-induced DNA interstrand crosslinks via the Nox-RIP1-ASK1-MAPK pathway
journal, May 2021
- Hirata, Yusuke; Takahashi, Miki; Yamada, Yuto
- Scientific Reports, Vol. 11, Issue 1
Genome engineering using the CRISPR-Cas9 system
journal, October 2013
- Ran, F. Ann; Hsu, Patrick D.; Wright, Jason
- Nature Protocols, Vol. 8, Issue 11
Mitochondrial One-Carbon Pathway Supports Cytosolic Folate Integrity in Cancer Cells
journal, November 2018
- Zheng, Yuxiang; Lin, Ting-Yu; Lee, Gina
- Cell, Vol. 175, Issue 6
Glucose-6-phosphate dehydrogenase deficiency
journal, January 2008
- Cappellini, Md; Fiorelli, G.
- The Lancet, Vol. 371, Issue 9606
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
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
Glucose-6-phosphate dehydrogenase deficiency.
journal, December 1995
- Kaplan, M.; Hammerman, C.
- Postgraduate Medical Journal, Vol. 71, Issue 842
Quantitative flux analysis reveals folate-dependent NADPH production
journal, May 2014
- Fan, Jing; Ye, Jiangbin; Kamphorst, Jurre J.
- Nature, Vol. 510, Issue 7504
Metabolite Spectral Accuracy on Orbitraps
journal, May 2017
- Su, Xiaoyang; Lu, Wenyun; Rabinowitz, Joshua D.
- Analytical Chemistry, Vol. 89, Issue 11
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
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
Tracing Compartmentalized NADPH Metabolism in the Cytosol and Mitochondria of Mammalian Cells
journal, July 2014
- Lewis, Caroline A.; Parker, Seth J.; Fiske, Brian P.
- Molecular Cell, Vol. 55, Issue 2
Serine and one-carbon metabolism in cancer
journal, September 2016
- Yang, Ming; Vousden, Karen H.
- Nature Reviews Cancer, Vol. 16, Issue 10
Kinetic mechanism of glucose-6-phosphate dehydrogenase from the lactating rat mammary gland. Implications for regulation.
journal, April 1980
- Shreve, D. S.; Levy, H. R.
- Journal of Biological Chemistry, Vol. 255, Issue 7
Unusual transient- and steady-state kinetic behavior is predicted by the kinetic scheme operational for recombinant human dihydrofolate reductase.
journal, February 1990
- Appleman, J. R.; Beard, W. A.; Delcamp, T. J.
- Journal of Biological Chemistry, Vol. 265, Issue 5
Studies on the Metabolism of Adipose Tissue
journal, March 1964
- Flatt, J. P.; Ball, Eric G.
- Journal of Biological Chemistry, Vol. 239, Issue 3
Works referencing / citing this record:
The PI3K–AKT network at the interface of oncogenic signalling and cancer metabolism
journal, November 2019
- Hoxhaj, Gerta; Manning, Brendan D.
- Nature Reviews Cancer, Vol. 20, Issue 2
Limitations of Deuterium-Labelled Substrates for Quantifying NADPH Metabolism in Heterotrophic Arabidopsis Cell Cultures
journal, September 2019
- Smith, Edward N.; McCullagh, James S. O.; Ratcliffe, R. George
- Metabolites, Vol. 9, Issue 10
The complexity of the serine glycine one-carbon pathway in cancer
journal, November 2019
- Reina-Campos, Miguel; Diaz-Meco, Maria T.; Moscat, Jorge
- Journal of Cell Biology, Vol. 219, Issue 1
Parabiosis Incompletely Reverses Aging-Induced Metabolic Changes and Oxidant Stress in Mouse Red Blood Cells
journal, June 2019
- Morrison, Evan J.; Champagne, Devin P.; Dzieciatkowska, Monika
- Nutrients, Vol. 11, Issue 6
The Redox Role of G6PD in Cell Growth, Cell Death, and Cancer
journal, September 2019
- Yang,
- Cells, Vol. 8, Issue 9
Parabiosis Incompletely Reverses Aging-Induced Metabolic Changes and Oxidant Stress in Mouse Red Blood Cells
journal, June 2019
- Morrison, Evan J.; Champagne, Devin P.; Dzieciatkowska, Monika
- Nutrients, Vol. 11, Issue 6