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GFPT2/GFAT2 and AMDHD2 act in tandem to control the hexosamine pathway

The hexosamine biosynthetic pathway (HBP) produces the essential metabolite UDP-GlcNAc and plays a key role in metabolism, health, and aging. The HBP is controlled by its rate-limiting enzyme glutamine fructose-6-phosphate amidotransferase (GFPT/GFAT) that is directly inhibited by UDP-GlcNAc in a fe...

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Autores principales: Kroef, Virginia, Ruegenberg, Sabine, Horn, Moritz, Allmeroth, Kira, Ebert, Lena, Bozkus, Seyma, Miethe, Stephan, Elling, Ulrich, Schermer, Bernhard, Baumann, Ulrich, Denzel, Martin Sebastian
Formato: Online Artículo Texto
Lenguaje:English
Publicado: eLife Sciences Publications, Ltd 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8970586/
https://www.ncbi.nlm.nih.gov/pubmed/35229715
http://dx.doi.org/10.7554/eLife.69223
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author Kroef, Virginia
Ruegenberg, Sabine
Horn, Moritz
Allmeroth, Kira
Ebert, Lena
Bozkus, Seyma
Miethe, Stephan
Elling, Ulrich
Schermer, Bernhard
Baumann, Ulrich
Denzel, Martin Sebastian
author_facet Kroef, Virginia
Ruegenberg, Sabine
Horn, Moritz
Allmeroth, Kira
Ebert, Lena
Bozkus, Seyma
Miethe, Stephan
Elling, Ulrich
Schermer, Bernhard
Baumann, Ulrich
Denzel, Martin Sebastian
author_sort Kroef, Virginia
collection PubMed
description The hexosamine biosynthetic pathway (HBP) produces the essential metabolite UDP-GlcNAc and plays a key role in metabolism, health, and aging. The HBP is controlled by its rate-limiting enzyme glutamine fructose-6-phosphate amidotransferase (GFPT/GFAT) that is directly inhibited by UDP-GlcNAc in a feedback loop. HBP regulation by GFPT is well studied but other HBP regulators have remained obscure. Elevated UDP-GlcNAc levels counteract the glycosylation toxin tunicamycin (TM), and thus we screened for TM resistance in haploid mouse embryonic stem cells (mESCs) using random chemical mutagenesis to determine alternative HBP regulation. We identified the N-acetylglucosamine deacetylase AMDHD2 that catalyzes a reverse reaction in the HBP and its loss strongly elevated UDP-GlcNAc. To better understand AMDHD2, we solved the crystal structure and found that loss-of-function (LOF) is caused by protein destabilization or interference with its catalytic activity. Finally, we show that mESCs express AMDHD2 together with GFPT2 instead of the more common paralog GFPT1. Compared with GFPT1, GFPT2 had a much lower sensitivity to UDP-GlcNAc inhibition, explaining how AMDHD2 LOF resulted in HBP activation. This HBP configuration in which AMDHD2 serves to balance GFPT2 activity was also observed in other mESCs and, consistently, the GFPT2:GFPT1 ratio decreased with differentiation of human embryonic stem cells. Taken together, our data reveal a critical function of AMDHD2 in limiting UDP-GlcNAc production in cells that use GFPT2 for metabolite entry into the HBP.
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spelling pubmed-89705862022-04-01 GFPT2/GFAT2 and AMDHD2 act in tandem to control the hexosamine pathway Kroef, Virginia Ruegenberg, Sabine Horn, Moritz Allmeroth, Kira Ebert, Lena Bozkus, Seyma Miethe, Stephan Elling, Ulrich Schermer, Bernhard Baumann, Ulrich Denzel, Martin Sebastian eLife Biochemistry and Chemical Biology The hexosamine biosynthetic pathway (HBP) produces the essential metabolite UDP-GlcNAc and plays a key role in metabolism, health, and aging. The HBP is controlled by its rate-limiting enzyme glutamine fructose-6-phosphate amidotransferase (GFPT/GFAT) that is directly inhibited by UDP-GlcNAc in a feedback loop. HBP regulation by GFPT is well studied but other HBP regulators have remained obscure. Elevated UDP-GlcNAc levels counteract the glycosylation toxin tunicamycin (TM), and thus we screened for TM resistance in haploid mouse embryonic stem cells (mESCs) using random chemical mutagenesis to determine alternative HBP regulation. We identified the N-acetylglucosamine deacetylase AMDHD2 that catalyzes a reverse reaction in the HBP and its loss strongly elevated UDP-GlcNAc. To better understand AMDHD2, we solved the crystal structure and found that loss-of-function (LOF) is caused by protein destabilization or interference with its catalytic activity. Finally, we show that mESCs express AMDHD2 together with GFPT2 instead of the more common paralog GFPT1. Compared with GFPT1, GFPT2 had a much lower sensitivity to UDP-GlcNAc inhibition, explaining how AMDHD2 LOF resulted in HBP activation. This HBP configuration in which AMDHD2 serves to balance GFPT2 activity was also observed in other mESCs and, consistently, the GFPT2:GFPT1 ratio decreased with differentiation of human embryonic stem cells. Taken together, our data reveal a critical function of AMDHD2 in limiting UDP-GlcNAc production in cells that use GFPT2 for metabolite entry into the HBP. eLife Sciences Publications, Ltd 2022-03-01 /pmc/articles/PMC8970586/ /pubmed/35229715 http://dx.doi.org/10.7554/eLife.69223 Text en © 2022, Kroef et al https://creativecommons.org/licenses/by/4.0/This article is distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use and redistribution provided that the original author and source are credited.
spellingShingle Biochemistry and Chemical Biology
Kroef, Virginia
Ruegenberg, Sabine
Horn, Moritz
Allmeroth, Kira
Ebert, Lena
Bozkus, Seyma
Miethe, Stephan
Elling, Ulrich
Schermer, Bernhard
Baumann, Ulrich
Denzel, Martin Sebastian
GFPT2/GFAT2 and AMDHD2 act in tandem to control the hexosamine pathway
title GFPT2/GFAT2 and AMDHD2 act in tandem to control the hexosamine pathway
title_full GFPT2/GFAT2 and AMDHD2 act in tandem to control the hexosamine pathway
title_fullStr GFPT2/GFAT2 and AMDHD2 act in tandem to control the hexosamine pathway
title_full_unstemmed GFPT2/GFAT2 and AMDHD2 act in tandem to control the hexosamine pathway
title_short GFPT2/GFAT2 and AMDHD2 act in tandem to control the hexosamine pathway
title_sort gfpt2/gfat2 and amdhd2 act in tandem to control the hexosamine pathway
topic Biochemistry and Chemical Biology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8970586/
https://www.ncbi.nlm.nih.gov/pubmed/35229715
http://dx.doi.org/10.7554/eLife.69223
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