Loss of GFAT-1 Feedback Regulation Activates the Hexosamine Pathway That Modulates Protein Homeostasis

Overview

Journal Nat Commun

Specialty Biology

Date 2020 Feb 6

PMID 32019926

Citations 27

Authors

Sabine Ruegenberg

Moritz Horn

Christian Pichlo

Kira Allmeroth

Ulrich Baumann

Martin S Denzel

Affiliations

Soon will be listed here.

Abstract

Glutamine fructose-6-phosphate amidotransferase (GFAT) is the key enzyme in the hexosamine pathway (HP) that produces uridine 5'-diphospho-N-acetyl-D-glucosamine (UDP-GlcNAc), linking energy metabolism with posttranslational protein glycosylation. In Caenorhabditis elegans, we previously identified gfat-1 gain-of-function mutations that elevate UDP-GlcNAc levels, improve protein homeostasis, and extend lifespan. GFAT is highly conserved, but the gain-of-function mechanism and its relevance in mammalian cells remained unclear. Here, we present the full-length crystal structure of human GFAT-1 in complex with various ligands and with important mutations. UDP-GlcNAc directly interacts with GFAT-1, inhibiting catalytic activity. The longevity-associated G451E variant shows drastically reduced sensitivity to UDP-GlcNAc inhibition in enzyme activity assays. Our structural and functional data point to a critical role of the interdomain linker in UDP-GlcNAc inhibition. In mammalian cells, the G451E variant potently activates the HP. Therefore, GFAT-1 gain-of-function through loss of feedback inhibition constitutes a potential target for the treatment of age-related proteinopathies.

Citing Articles

Gfa1 (glutamine fructose-6-phosphate aminotransferase) is essential for Aspergillus fumigatus growth and virulence.

Qin Q, Wei P, Usman S, Ahamefule C, Jin C, Wang B BMC Biol. 2025; 23(1):80.

PMID: 40082985 PMC: 11907850. DOI: 10.1186/s12915-025-02184-0.

Crosstalk between O-GlcNAcylation and phosphorylation in metabolism: regulation and mechanism.

Zhao Q, Zhou S, Lou W, Qian H, Xu Z Cell Death Differ. 2025; .

PMID: 40045035 DOI: 10.1038/s41418-025-01473-z.

An Optimized Marinopyrrole A Derivative Targets 6-Phosphoglucosamine Synthetase to Inhibit Methicillin-Resistant .

Guo F, Xiao F, Song H, Li X, Xiao Y, Qin Y ACS Cent Sci. 2024; 10(11):2090-2098.

PMID: 39634224 PMC: 11613329. DOI: 10.1021/acscentsci.4c01167.

GFPT2: A novel biomarker in mesothelioma for diagnosis and prognosis and its molecular mechanism in malignant progression.

Wei J, Zhou S, Chen G, Chen T, Wang Y, Zou J Br J Cancer. 2024; 131(9):1529-1542.

PMID: 39317702 PMC: 11519369. DOI: 10.1038/s41416-024-02830-4.

O-GlcNAcylation mediates Wnt-stimulated bone formation by rewiring aerobic glycolysis.

You C, Shen F, Yang P, Cui J, Ren Q, Liu M EMBO Rep. 2024; 25(10):4465-4487.

PMID: 39256595 PMC: 11467389. DOI: 10.1038/s44319-024-00237-z.

References

Kenyon C . The genetics of ageing. Nature. 2010; 464(7288):504-12. DOI: 10.1038/nature08980. View

Mannick J, Morris M, Hockey H, Roma G, Beibel M, Kulmatycki K . TORC1 inhibition enhances immune function and reduces infections in the elderly. Sci Transl Med. 2018; 10(449). DOI: 10.1126/scitranslmed.aaq1564. View

Newman J, Milman S, Hashmi S, Austad S, Kirkland J, Halter J . Strategies and Challenges in Clinical Trials Targeting Human Aging. J Gerontol A Biol Sci Med Sci. 2016; 71(11):1424-1434. PMC: 5055653. DOI: 10.1093/gerona/glw149. View

Denzel M, Storm N, Gutschmidt A, Baddi R, Hinze Y, Jarosch E . Hexosamine pathway metabolites enhance protein quality control and prolong life. Cell. 2014; 156(6):1167-1178. DOI: 10.1016/j.cell.2014.01.061. View

Marshall S, Bacote V, Traxinger R . Discovery of a metabolic pathway mediating glucose-induced desensitization of the glucose transport system. Role of hexosamine biosynthesis in the induction of insulin resistance. J Biol Chem. 1991; 266(8):4706-12. View

Parodi A . Role of N-oligosaccharide endoplasmic reticulum processing reactions in glycoprotein folding and degradation. Biochem J. 2000; 348 Pt 1:1-13. PMC: 1221029. View

Hanisch F . O-glycosylation of the mucin type. Biol Chem. 2001; 382(2):143-9. DOI: 10.1515/BC.2001.022. View

Hart G . Dynamic O-linked glycosylation of nuclear and cytoskeletal proteins. Annu Rev Biochem. 1997; 66:315-35. DOI: 10.1146/annurev.biochem.66.1.315. View

Yang X, Qian K . Protein O-GlcNAcylation: emerging mechanisms and functions. Nat Rev Mol Cell Biol. 2017; 18(7):452-465. PMC: 5667541. DOI: 10.1038/nrm.2017.22. View

10.

van der Laarse S, Leney A, Heck A . Crosstalk between phosphorylation and O-GlcNAcylation: friend or foe. FEBS J. 2018; 285(17):3152-3167. DOI: 10.1111/febs.14491. View

11.

Gong C, Liu F, Iqbal K . O-GlcNAcylation: A regulator of tau pathology and neurodegeneration. Alzheimers Dement. 2016; 12(10):1078-1089. DOI: 10.1016/j.jalz.2016.02.011. View

12.

Liu F, Iqbal K, Grundke-Iqbal I, Hart G, Gong C . O-GlcNAcylation regulates phosphorylation of tau: a mechanism involved in Alzheimer's disease. Proc Natl Acad Sci U S A. 2004; 101(29):10804-9. PMC: 490015. DOI: 10.1073/pnas.0400348101. View

13.

Hastings N, Wang X, Song L, Butts B, Grotz D, Hargreaves R . Inhibition of O-GlcNAcase leads to elevation of O-GlcNAc tau and reduction of tauopathy and cerebrospinal fluid tau in rTg4510 mice. Mol Neurodegener. 2017; 12(1):39. PMC: 5437664. DOI: 10.1186/s13024-017-0181-0. View

14.

Wang Z, Deng Y, Gao N, Pedrozo Z, Li D, Morales C . Spliced X-box binding protein 1 couples the unfolded protein response to hexosamine biosynthetic pathway. Cell. 2014; 156(6):1179-1192. PMC: 3959665. DOI: 10.1016/j.cell.2014.01.014. View

15.

Ghosh S, BLUMENTHAL H, Davidson E, Roseman S . Glucosamine metabolism. V. Enzymatic synthesis of glucosamine 6-phosphate. J Biol Chem. 1960; 235:1265-73. View

16.

Denisot M, Le Goffic F, Badet B . Glucosamine-6-phosphate synthase from Escherichia coli yields two proteins upon limited proteolysis: identification of the glutamine amidohydrolase and 2R ketose/aldose isomerase-bearing domains based on their biochemical properties. Arch Biochem Biophys. 1991; 288(1):225-30. DOI: 10.1016/0003-9861(91)90188-o. View

17.

Mouilleron S, Badet-Denisot M, Golinelli-Pimpaneau B . Glutamine binding opens the ammonia channel and activates glucosamine-6P synthase. J Biol Chem. 2005; 281(7):4404-12. DOI: 10.1074/jbc.M511689200. View

18.

KORNFELD R . Studies on L-glutamine D-fructose 6-phosphate amidotransferase. I. Feedback inhibition by uridine diphosphate-N-acetylglucosamine. J Biol Chem. 1967; 242(13):3135-41. View

19.

Vessal M, HASSID W . Partial Purification and Properties of l-Glutamine d-Fructose 6-Phosphate Amidotransferase from Phaseolus aureus. Plant Physiol. 1972; 49(6):977-81. PMC: 366091. DOI: 10.1104/pp.49.6.977. View

20.

Milewski S, Kuszczak D, Jedrzejczak R, Smith R, Brown A, Gooday G . Oligomeric structure and regulation of Candida albicans glucosamine-6-phosphate synthase. J Biol Chem. 1999; 274(7):4000-8. DOI: 10.1074/jbc.274.7.4000. View