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Formate-tetrahydrofolate ligase: supplying the cytosolic one-carbon network in roots with one-carbon units originating from glycolate.
Saeheng, Sompop; Bailes, Clayton; Bao, Han; Gashu, Kelem; Morency, Matt; Arlynn, Tana; Smertenko, Andrei; Walker, Berkley James; Roje, Sanja.
Afiliação
  • Saeheng S; Institute of Biological Chemistry, Washington State University, Pullman, Washington, USA.
  • Bailes C; Center of Excellence for Biochemistry, Faculty of Science, Prince of Songkla University, Hat Yai, 90110, Thailand.
  • Bao H; Plant Cell and Physiology for Sustainable Agriculture Research Unit, Faculty of Science, Prince of Songkla University, Hat Yai, 90110, Thailand.
  • Gashu K; Institute of Biological Chemistry, Washington State University, Pullman, Washington, USA.
  • Morency M; Department of Energy-Michigan State University Plant Research Laboratory, Michigan State University, East Lansing, Michigan, USA.
  • Arlynn T; Department of Energy-Michigan State University Plant Research Laboratory, Michigan State University, East Lansing, Michigan, USA.
  • Smertenko A; Department of Plant Biology, Michigan State University, East Lansing, Michigan, USA.
  • Walker BJ; Department of Energy-Michigan State University Plant Research Laboratory, Michigan State University, East Lansing, Michigan, USA.
  • Roje S; Department of Plant Biology, Michigan State University, East Lansing, Michigan, USA.
Plant J ; 2024 Jul 16.
Article em En | MEDLINE | ID: mdl-39010784
ABSTRACT
The metabolism of tetrahydrofolate (H4PteGlun)-bound one-carbon (C1) units (C1 metabolism) is multifaceted and required for plant growth, but it is unclear what of many possible synthesis pathways provide C1 units in specific organelles and tissues. One possible source of C1 units is via formate-tetrahydrofolate ligase, which catalyzes the reversible ATP-driven production of 10-formyltetrahydrofolate (10-formyl-H4PteGlun) from formate and tetrahydrofolate (H4PteGlun). Here, we report biochemical and functional characterization of the enzyme from Arabidopsis thaliana (AtFTHFL). We show that the recombinant AtFTHFL has lower Km and kcat values with pentaglutamyl tetrahydrofolate (H4PteGlu5) as compared to monoglutamyl tetrahydrofolate (H4PteGlu1), 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 H4PteGlun + 5,10-methylene-H4PteGlun 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 C1 network in roots with C1 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-H4PteGlun, 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.
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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Idioma: En Revista: Plant J Assunto da revista: BIOLOGIA MOLECULAR / BOTANICA Ano de publicação: 2024 Tipo de documento: Article País de afiliação: Estados Unidos

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Idioma: En Revista: Plant J Assunto da revista: BIOLOGIA MOLECULAR / BOTANICA Ano de publicação: 2024 Tipo de documento: Article País de afiliação: Estados Unidos