Exploratory Metabolomics Underscores the Folate Enzyme ALDH1L1 as a Regulator of Glycine and Methylation Reactions
Folate (vitamin B9) is involved in one-carbon transfer reactions and plays a significant role in nucleic acid synthesis and control of cellular proliferation, among other key cellular processes. It is now recognized that the role of folates in different stages of carcinogenesis is complex, and more...
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| Format: | Article |
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MDPI AG
2022-12-01
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| Series: | Molecules |
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| Online Access: | https://www.mdpi.com/1420-3049/27/23/8394 |
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| author | Blake R. Rushing Halle M. Fogle Jaspreet Sharma Mikyoung You Jonathan P. McCormac Sabrina Molina Susan Sumner Natalia I. Krupenko Sergey A. Krupenko |
| author_facet | Blake R. Rushing Halle M. Fogle Jaspreet Sharma Mikyoung You Jonathan P. McCormac Sabrina Molina Susan Sumner Natalia I. Krupenko Sergey A. Krupenko |
| author_sort | Blake R. Rushing |
| collection | DOAJ |
| description | Folate (vitamin B9) is involved in one-carbon transfer reactions and plays a significant role in nucleic acid synthesis and control of cellular proliferation, among other key cellular processes. It is now recognized that the role of folates in different stages of carcinogenesis is complex, and more research is needed to understand how folate reactions become dysregulated in cancers and the metabolic consequences that occur as a result. ALDH1L1 (cytosolic 10-formyltetrahydrofolate dehydrogenase), an enzyme of folate metabolism expressed in many tissues, is ubiquitously downregulated in cancers and is not expressed in cancer cell lines. The RT4 cell line (derived from papillary bladder cancer) which expresses high levels of ALDH1L1 represents an exception, providing an opportunity to explore the metabolic consequences of the loss of this enzyme. We have downregulated this protein in RT4 cells (shRNA driven knockdown or CRISPR driven knockout) and compared metabolomes of ALDH1L1-expressing and -deficient cells to determine if metabolic changes linked to the loss of this enzyme might provide proliferative and/or survival advantages for cancer cells. In this study, cell extracts were analyzed using Ultra High Performance Liquid Chromatography High Resolution Mass Spectrometry (UHPLC-HR-MS). A total of 13,339 signals were identified or annotated using an in-house library and public databases. Supervised and unsupervised multivariate analysis revealed metabolic differences between RT4 cells and ALDH1L1-deficient clones. Glycine (8-fold decrease) and metabolites derived from S-adenosylmethionine utilizing pathways were significantly decreased in the ALDH1L1-deficient clones, compared with RT4 cells. Other changes linked to ALDH1L1 downregulation include decreased levels of amino acids, Krebs cycle intermediates, and ribose-5-phosphate, and increased nicotinic acid. While the ALDH1L1-catalyzed reaction is directly linked to glycine biosynthesis and methyl group flux, its overall effect on cellular metabolism extends beyond immediate metabolic pathways controlled by this enzyme. |
| first_indexed | 2024-03-09T17:40:00Z |
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| id | doaj.art-c236192cec5040e59ea682b9f035d9a0 |
| institution | Directory Open Access Journal |
| issn | 1420-3049 |
| language | English |
| last_indexed | 2024-03-09T17:40:00Z |
| publishDate | 2022-12-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Molecules |
| spelling | doaj.art-c236192cec5040e59ea682b9f035d9a02023-11-24T11:41:28ZengMDPI AGMolecules1420-30492022-12-012723839410.3390/molecules27238394Exploratory Metabolomics Underscores the Folate Enzyme ALDH1L1 as a Regulator of Glycine and Methylation ReactionsBlake R. Rushing0Halle M. Fogle1Jaspreet Sharma2Mikyoung You3Jonathan P. McCormac4Sabrina Molina5Susan Sumner6Natalia I. Krupenko7Sergey A. Krupenko8Nutrition Research Institute, UNC Chapel Hill, Kannapolis, NC 28081, USANutrition Research Institute, UNC Chapel Hill, Kannapolis, NC 28081, USANutrition Research Institute, UNC Chapel Hill, Kannapolis, NC 28081, USANutrition Research Institute, UNC Chapel Hill, Kannapolis, NC 28081, USANutrition Research Institute, UNC Chapel Hill, Kannapolis, NC 28081, USANutrition Research Institute, UNC Chapel Hill, Kannapolis, NC 28081, USANutrition Research Institute, UNC Chapel Hill, Kannapolis, NC 28081, USANutrition Research Institute, UNC Chapel Hill, Kannapolis, NC 28081, USANutrition Research Institute, UNC Chapel Hill, Kannapolis, NC 28081, USAFolate (vitamin B9) is involved in one-carbon transfer reactions and plays a significant role in nucleic acid synthesis and control of cellular proliferation, among other key cellular processes. It is now recognized that the role of folates in different stages of carcinogenesis is complex, and more research is needed to understand how folate reactions become dysregulated in cancers and the metabolic consequences that occur as a result. ALDH1L1 (cytosolic 10-formyltetrahydrofolate dehydrogenase), an enzyme of folate metabolism expressed in many tissues, is ubiquitously downregulated in cancers and is not expressed in cancer cell lines. The RT4 cell line (derived from papillary bladder cancer) which expresses high levels of ALDH1L1 represents an exception, providing an opportunity to explore the metabolic consequences of the loss of this enzyme. We have downregulated this protein in RT4 cells (shRNA driven knockdown or CRISPR driven knockout) and compared metabolomes of ALDH1L1-expressing and -deficient cells to determine if metabolic changes linked to the loss of this enzyme might provide proliferative and/or survival advantages for cancer cells. In this study, cell extracts were analyzed using Ultra High Performance Liquid Chromatography High Resolution Mass Spectrometry (UHPLC-HR-MS). A total of 13,339 signals were identified or annotated using an in-house library and public databases. Supervised and unsupervised multivariate analysis revealed metabolic differences between RT4 cells and ALDH1L1-deficient clones. Glycine (8-fold decrease) and metabolites derived from S-adenosylmethionine utilizing pathways were significantly decreased in the ALDH1L1-deficient clones, compared with RT4 cells. Other changes linked to ALDH1L1 downregulation include decreased levels of amino acids, Krebs cycle intermediates, and ribose-5-phosphate, and increased nicotinic acid. While the ALDH1L1-catalyzed reaction is directly linked to glycine biosynthesis and methyl group flux, its overall effect on cellular metabolism extends beyond immediate metabolic pathways controlled by this enzyme.https://www.mdpi.com/1420-3049/27/23/8394folateALDH1L1cancermetabolomics |
| spellingShingle | Blake R. Rushing Halle M. Fogle Jaspreet Sharma Mikyoung You Jonathan P. McCormac Sabrina Molina Susan Sumner Natalia I. Krupenko Sergey A. Krupenko Exploratory Metabolomics Underscores the Folate Enzyme ALDH1L1 as a Regulator of Glycine and Methylation Reactions Molecules folate ALDH1L1 cancer metabolomics |
| title | Exploratory Metabolomics Underscores the Folate Enzyme ALDH1L1 as a Regulator of Glycine and Methylation Reactions |
| title_full | Exploratory Metabolomics Underscores the Folate Enzyme ALDH1L1 as a Regulator of Glycine and Methylation Reactions |
| title_fullStr | Exploratory Metabolomics Underscores the Folate Enzyme ALDH1L1 as a Regulator of Glycine and Methylation Reactions |
| title_full_unstemmed | Exploratory Metabolomics Underscores the Folate Enzyme ALDH1L1 as a Regulator of Glycine and Methylation Reactions |
| title_short | Exploratory Metabolomics Underscores the Folate Enzyme ALDH1L1 as a Regulator of Glycine and Methylation Reactions |
| title_sort | exploratory metabolomics underscores the folate enzyme aldh1l1 as a regulator of glycine and methylation reactions |
| topic | folate ALDH1L1 cancer metabolomics |
| url | https://www.mdpi.com/1420-3049/27/23/8394 |
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