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The transfer of one-carbon (C1) units is an integral part of cellular metabolism and is essential for the biosynthesis of nucleotides-, amino acids, and cofactors, as well as for cellular methylation reactions. Within the plant cell, mitochondria are considered to be the hub of one-carbon metabolism, but the mechanisms and fluxes that distribute C1 units from the mitochondria throughout the cell are unknown. Formate, the anion of formic acid, is an intermediate of C1 metabolism and is converted to C1-tetrahydrofolate intermediates (C1 folates) or oxidized to CO2 by formate dehydrogenase. The existence of formate dehydrogenase in plant cells challenges the formate exchange between mitochondria and the cytosol, a basic principle of eukaryotic cellular and organellar C1 metabolism. Based on the biochemical and physiological characterization of Arabidopsis thaliana formate dehydrogenase 1 (FDH1), we propose an FDH1-regulated C1 shunt linking mitochondrial and cytosolic C1 metabolism by formate exchange. Finally, we give a perspective on a cellular serine/formate shuttle that allows the distribution and transfer of C1 units according to the redox state within the compartments.
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An enzymatic activation of formaldehyde for nucleotide methylation
Abstract Folate enzyme cofactors and their derivatives have the unique ability to provide a single carbon unit at different oxidation levels for the de novo synthesis of amino-acids, purines, or thymidylate, an essential DNA nucleotide. How these cofactors mediate methylene transfer is not fully settled yet, particularly with regard to how the methylene is transferred to the methylene acceptor. Here, we uncovered that the bacterial thymidylate synthase ThyX, which relies on both folate and flavin for activity, can also use a formaldehyde-shunt to directly synthesize thymidylate. Combining biochemical, spectroscopic and anaerobic crystallographic analyses, we showed that formaldehyde reacts with the reduced flavin coenzyme to form a carbinolamine intermediate used by ThyX for dUMP methylation. The crystallographic structure of this intermediate reveals how ThyX activates formaldehyde and uses it, with the assistance of active site residues, to methylate dUMP. Our results reveal that carbinolamine species promote methylene transfer and suggest that the use of a CH 2 O-shunt may be relevant in several other important folate-dependent reactions.
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- Award ID(s):
- 1905267
- PAR ID:
- 10310052
- Date Published:
- Journal Name:
- Nature Communications
- Volume:
- 12
- Issue:
- 1
- ISSN:
- 2041-1723
- Format(s):
- Medium: X
- Sponsoring Org:
- National Science Foundation
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- Free Publicly Accessible Full Text
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Accepted Manuscript1.0
- Journal Article:
- https://doi.org/10.1038/s41467-021-24756-8
