Mthfs is an Essential Gene in Mice and a Component of the Purinosome
Tetrahydrofolates (THF) are a family of cofactors that function as one-carbon donors in folate-dependent one-carbon metabolism, a metabolic network required for the de novo synthesis of purines, thymidylate, and for the remethylation of homocysteine to methionine in the cytoplasm. 5-FormylTHF is not...
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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Frontiers Research Foundation
2011
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3268590/ https://www.ncbi.nlm.nih.gov/pubmed/22303332 http://dx.doi.org/10.3389/fgene.2011.00036 |
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author | Field, Martha S. Anderson, Donald D. Stover, Patrick J. |
author_facet | Field, Martha S. Anderson, Donald D. Stover, Patrick J. |
author_sort | Field, Martha S. |
collection | PubMed |
description | Tetrahydrofolates (THF) are a family of cofactors that function as one-carbon donors in folate-dependent one-carbon metabolism, a metabolic network required for the de novo synthesis of purines, thymidylate, and for the remethylation of homocysteine to methionine in the cytoplasm. 5-FormylTHF is not a cofactor in one-carbon metabolism, but serves as a storage form of THF cofactors. 5-formylTHF is mobilized back into the THF cofactor pool by methenyltetrahydrofolate synthetase (MTHFS), which catalyzes the irreversible and ATP-dependent conversion 5-formyltetrahydrofolate to 5,10-methenyltetrahydrofolate. Mthfs is not an essential gene in Arabidopsis, but MTHFS expression is elevated in animal tumors, enhances de novo purine synthesis, confers partial resistance to antifolate purine synthesis inhibitors and increases rates of folate catabolism in mammalian cell cultures. However, the mechanisms underlying these effects of MTHFS expression have yet to be established. The purpose of this study was to investigate the role and essentiality of MTHFS in mice. Mthfs was disrupted through the insertion of a gene trap vector between exons 1 and 2. Mthfs(gt/+) mice were fertile and viable. No Mthfs(gt/gt) embryos were recovered from Mthfs(gt/+) intercrosses, indicating Mthfs is an essential gene in mice. Tissue MTHFS protein levels are decreased in both Mthfs(gt/+) and Mthfs(+/+) mice placed on a folate and choline deficient diet, and mouse embryonic fibroblasts from Mthfs(gt/+) embryos exhibit decreased capacity for de novo purine synthesis without impairment in de novo thymidylate synthesis. MTHFS was shown to co-localize with two enzymes of the de novo purine synthesis pathway in HeLa cells in a cell cycle-dependent manner, and to be modified by the small ubiquitin-like modifier (SUMO) protein. Mutation of the consensus SUMO modification sites on MTHFS eliminated co-localization of MTHFS with the de novo purine biosynthesis pathway under purine-deficient conditions. The results from this study indicate that MTHFS enhances purine biosynthesis by delivering 10-formylTHF to the purinosome in a SUMO-dependent fashion. |
format | Online Article Text |
id | pubmed-3268590 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2011 |
publisher | Frontiers Research Foundation |
record_format | MEDLINE/PubMed |
spelling | pubmed-32685902012-02-02 Mthfs is an Essential Gene in Mice and a Component of the Purinosome Field, Martha S. Anderson, Donald D. Stover, Patrick J. Front Genet Genetics Tetrahydrofolates (THF) are a family of cofactors that function as one-carbon donors in folate-dependent one-carbon metabolism, a metabolic network required for the de novo synthesis of purines, thymidylate, and for the remethylation of homocysteine to methionine in the cytoplasm. 5-FormylTHF is not a cofactor in one-carbon metabolism, but serves as a storage form of THF cofactors. 5-formylTHF is mobilized back into the THF cofactor pool by methenyltetrahydrofolate synthetase (MTHFS), which catalyzes the irreversible and ATP-dependent conversion 5-formyltetrahydrofolate to 5,10-methenyltetrahydrofolate. Mthfs is not an essential gene in Arabidopsis, but MTHFS expression is elevated in animal tumors, enhances de novo purine synthesis, confers partial resistance to antifolate purine synthesis inhibitors and increases rates of folate catabolism in mammalian cell cultures. However, the mechanisms underlying these effects of MTHFS expression have yet to be established. The purpose of this study was to investigate the role and essentiality of MTHFS in mice. Mthfs was disrupted through the insertion of a gene trap vector between exons 1 and 2. Mthfs(gt/+) mice were fertile and viable. No Mthfs(gt/gt) embryos were recovered from Mthfs(gt/+) intercrosses, indicating Mthfs is an essential gene in mice. Tissue MTHFS protein levels are decreased in both Mthfs(gt/+) and Mthfs(+/+) mice placed on a folate and choline deficient diet, and mouse embryonic fibroblasts from Mthfs(gt/+) embryos exhibit decreased capacity for de novo purine synthesis without impairment in de novo thymidylate synthesis. MTHFS was shown to co-localize with two enzymes of the de novo purine synthesis pathway in HeLa cells in a cell cycle-dependent manner, and to be modified by the small ubiquitin-like modifier (SUMO) protein. Mutation of the consensus SUMO modification sites on MTHFS eliminated co-localization of MTHFS with the de novo purine biosynthesis pathway under purine-deficient conditions. The results from this study indicate that MTHFS enhances purine biosynthesis by delivering 10-formylTHF to the purinosome in a SUMO-dependent fashion. Frontiers Research Foundation 2011-06-20 /pmc/articles/PMC3268590/ /pubmed/22303332 http://dx.doi.org/10.3389/fgene.2011.00036 Text en Copyright © 2011 Field, Anderson and Stover. http://www.frontiersin.org/licenseagreement This is an open-access article subject to a non-exclusive license between the authors and Frontiers Media SA, which permits use, distribution and reproduction in other forums, provided the original authors and source are credited and other Frontiers conditions are complied with. |
spellingShingle | Genetics Field, Martha S. Anderson, Donald D. Stover, Patrick J. Mthfs is an Essential Gene in Mice and a Component of the Purinosome |
title | Mthfs is an Essential Gene in Mice and a Component of the Purinosome |
title_full | Mthfs is an Essential Gene in Mice and a Component of the Purinosome |
title_fullStr | Mthfs is an Essential Gene in Mice and a Component of the Purinosome |
title_full_unstemmed | Mthfs is an Essential Gene in Mice and a Component of the Purinosome |
title_short | Mthfs is an Essential Gene in Mice and a Component of the Purinosome |
title_sort | mthfs is an essential gene in mice and a component of the purinosome |
topic | Genetics |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3268590/ https://www.ncbi.nlm.nih.gov/pubmed/22303332 http://dx.doi.org/10.3389/fgene.2011.00036 |
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