Formate-tetrahydrofolate ligase: supplying the cytosolic one-carbon network in roots with one-carbon units originating from glycolate.

Autor: Saeheng S; Institute of Biological Chemistry, Washington State University, Pullman, Washington, USA.; Center of Excellence for Biochemistry, Faculty of Science, Prince of Songkla University, Hat Yai, 90110, Thailand.; Plant Cell and Physiology for Sustainable Agriculture Research Unit, Faculty of Science, Prince of Songkla University, Hat Yai, 90110, Thailand., Bailes C; Institute of Biological Chemistry, Washington State University, Pullman, Washington, USA., Bao H; Department of Energy-Michigan State University Plant Research Laboratory, Michigan State University, East Lansing, Michigan, USA., Gashu K; Department of Energy-Michigan State University Plant Research Laboratory, Michigan State University, East Lansing, Michigan, USA.; Department of Plant Biology, Michigan State University, East Lansing, Michigan, USA., Morency M; Department of Energy-Michigan State University Plant Research Laboratory, Michigan State University, East Lansing, Michigan, USA.; Department of Plant Biology, Michigan State University, East Lansing, Michigan, USA., Arlynn T; Institute of Biological Chemistry, Washington State University, Pullman, Washington, USA., Smertenko A; Institute of Biological Chemistry, Washington State University, Pullman, Washington, USA., Walker BJ; Department of Energy-Michigan State University Plant Research Laboratory, Michigan State University, East Lansing, Michigan, USA.; Department of Plant Biology, Michigan State University, East Lansing, Michigan, USA., Roje S; Institute of Biological Chemistry, Washington State University, Pullman, Washington, USA.
Jazyk: angličtina
Zdroj: The Plant journal : for cell and molecular biology [Plant J] 2024 Sep; Vol. 119 (5), pp. 2464-2483. Date of Electronic Publication: 2024 Jul 16.
DOI: 10.1111/tpj.16933
Abstrakt: The metabolism of tetrahydrofolate (H 4 PteGlu n )-bound one-carbon (C 1 ) units (C 1 metabolism) is multifaceted and required for plant growth, but it is unclear what of many possible synthesis pathways provide C 1 units in specific organelles and tissues. One possible source of C 1 units is via formate-tetrahydrofolate ligase, which catalyzes the reversible ATP-driven production of 10-formyltetrahydrofolate (10-formyl-H 4 PteGlu n ) from formate and tetrahydrofolate (H 4 PteGlu n ). Here, we report biochemical and functional characterization of the enzyme from Arabidopsis thaliana (AtFTHFL). We show that the recombinant AtFTHFL has lower K m and k cat values with pentaglutamyl tetrahydrofolate (H 4 PteGlu 5 ) as compared to monoglutamyl tetrahydrofolate (H 4 PteGlu 1 ), resulting in virtually identical catalytic efficiencies for the two substrates. Stable transformation of Arabidopsis plants with the EGFP-tagged AtFTHFL, followed with fluorescence microscopy, demonstrated cytosolic signal. Two independent T-DNA insertion lines with impaired AtFTHFL function had shorter roots compared to the wild type plants, demonstrating the importance of this enzyme for root growth. Overexpressing AtFTHFL led to the accumulation of H 4 PteGlu n  + 5,10-methylene-H 4 PteGlu n and serine, accompanied with the depletion of formate and glycolate, in roots of the transgenic Arabidopsis plants. This metabolic adjustment supports the hypothesis that AtFTHFL feeds the cytosolic C 1 network in roots with C 1 units originating from glycolate, and that these units are then used mainly for biosynthesis of serine, and not as much for the biosynthesis of 5-methyl-H 4 PteGlu n , methionine, and S-adenosylmethionine. This finding has implications for any future attempts to engineer one-carbon unit-requiring products through manipulation of the one-carbon metabolic network in non-photosynthetic organs.
(© 2024 The Author(s). The Plant Journal published by Society for Experimental Biology and John Wiley & Sons Ltd.)
Databáze: MEDLINE
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