Arabidopsis 3‐Deoxy‐d‐Arabino‐Heptulosonate 7‐Phosphate (DAHP) Synthases of the Shikimate Pathway Display Both Manganese‐ and Cobalt‐Dependent Activities
ABSTRACT The plant shikimate pathway directs a significant portion of photosynthetically assimilated carbon into the downstream biosynthetic pathways of aromatic amino acids (AAA) and aromatic natural products. 3‐Deoxy‐d‐arabino‐heptulosonate 7‐phosphate (DAHP) synthase (hereafter DHS) catalyzes the...
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| Language: | English |
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Wiley
2025-01-01
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| Series: | Plant Direct |
| Online Access: | https://doi.org/10.1002/pld3.70037 |
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| author | Ryo Yokoyama Hiroshi A. Maeda |
| author_facet | Ryo Yokoyama Hiroshi A. Maeda |
| author_sort | Ryo Yokoyama |
| collection | DOAJ |
| description | ABSTRACT The plant shikimate pathway directs a significant portion of photosynthetically assimilated carbon into the downstream biosynthetic pathways of aromatic amino acids (AAA) and aromatic natural products. 3‐Deoxy‐d‐arabino‐heptulosonate 7‐phosphate (DAHP) synthase (hereafter DHS) catalyzes the first step of the shikimate pathway, playing a critical role in controlling the carbon flux from central carbon metabolism into the AAA biosynthesis. Previous biochemical studies suggested the presence of manganese‐ and cobalt‐dependent DHS enzymes (DHS‐Mn and DHS‐Co, respectively) in various plant species. Unlike well‐studied DHS‐Mn, however, the identity of DHS‐Co is still unknown. Here, we show that all three DHS isoforms of Arabidopsis thaliana exhibit both DHS‐Mn and DHS‐Co activities in vitro. A phylogenetic analysis of various DHS orthologs and related sequences showed that Arabidopsis 3‐deoxy‐D‐manno‐octulosonate‐8‐phosphate synthase (KDOPS) proteins were closely related to microbial Type I DHSs. Despite their sequence similarity, these Arabidopsis KDOPS proteins showed no DHS activity. Meanwhile, optimization of the DHS assay conditions led to the successful detection of DHS‐Co activity from Arabidopsis DHS recombinant proteins. Compared with DHS‐Mn, DHS‐Co activity displayed the same redox dependency but distinct optimal pH and cofactor sensitivity. Our work provides biochemical evidence that the DHS isoforms of Arabidopsis possess DHS‐Co activity. |
| format | Article |
| id | doaj-art-d817c095587c40caaaae3c0d60b1c65e |
| institution | Kabale University |
| issn | 2475-4455 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | Wiley |
| record_format | Article |
| series | Plant Direct |
| spelling | doaj-art-d817c095587c40caaaae3c0d60b1c65e2025-01-30T04:44:29ZengWileyPlant Direct2475-44552025-01-0191n/an/a10.1002/pld3.70037Arabidopsis 3‐Deoxy‐d‐Arabino‐Heptulosonate 7‐Phosphate (DAHP) Synthases of the Shikimate Pathway Display Both Manganese‐ and Cobalt‐Dependent ActivitiesRyo Yokoyama0Hiroshi A. Maeda1Department of Botany University of Wisconsin‐Madison Madison Wisconsin USADepartment of Botany University of Wisconsin‐Madison Madison Wisconsin USAABSTRACT The plant shikimate pathway directs a significant portion of photosynthetically assimilated carbon into the downstream biosynthetic pathways of aromatic amino acids (AAA) and aromatic natural products. 3‐Deoxy‐d‐arabino‐heptulosonate 7‐phosphate (DAHP) synthase (hereafter DHS) catalyzes the first step of the shikimate pathway, playing a critical role in controlling the carbon flux from central carbon metabolism into the AAA biosynthesis. Previous biochemical studies suggested the presence of manganese‐ and cobalt‐dependent DHS enzymes (DHS‐Mn and DHS‐Co, respectively) in various plant species. Unlike well‐studied DHS‐Mn, however, the identity of DHS‐Co is still unknown. Here, we show that all three DHS isoforms of Arabidopsis thaliana exhibit both DHS‐Mn and DHS‐Co activities in vitro. A phylogenetic analysis of various DHS orthologs and related sequences showed that Arabidopsis 3‐deoxy‐D‐manno‐octulosonate‐8‐phosphate synthase (KDOPS) proteins were closely related to microbial Type I DHSs. Despite their sequence similarity, these Arabidopsis KDOPS proteins showed no DHS activity. Meanwhile, optimization of the DHS assay conditions led to the successful detection of DHS‐Co activity from Arabidopsis DHS recombinant proteins. Compared with DHS‐Mn, DHS‐Co activity displayed the same redox dependency but distinct optimal pH and cofactor sensitivity. Our work provides biochemical evidence that the DHS isoforms of Arabidopsis possess DHS‐Co activity.https://doi.org/10.1002/pld3.70037 |
| spellingShingle | Ryo Yokoyama Hiroshi A. Maeda Arabidopsis 3‐Deoxy‐d‐Arabino‐Heptulosonate 7‐Phosphate (DAHP) Synthases of the Shikimate Pathway Display Both Manganese‐ and Cobalt‐Dependent Activities Plant Direct |
| title | Arabidopsis 3‐Deoxy‐d‐Arabino‐Heptulosonate 7‐Phosphate (DAHP) Synthases of the Shikimate Pathway Display Both Manganese‐ and Cobalt‐Dependent Activities |
| title_full | Arabidopsis 3‐Deoxy‐d‐Arabino‐Heptulosonate 7‐Phosphate (DAHP) Synthases of the Shikimate Pathway Display Both Manganese‐ and Cobalt‐Dependent Activities |
| title_fullStr | Arabidopsis 3‐Deoxy‐d‐Arabino‐Heptulosonate 7‐Phosphate (DAHP) Synthases of the Shikimate Pathway Display Both Manganese‐ and Cobalt‐Dependent Activities |
| title_full_unstemmed | Arabidopsis 3‐Deoxy‐d‐Arabino‐Heptulosonate 7‐Phosphate (DAHP) Synthases of the Shikimate Pathway Display Both Manganese‐ and Cobalt‐Dependent Activities |
| title_short | Arabidopsis 3‐Deoxy‐d‐Arabino‐Heptulosonate 7‐Phosphate (DAHP) Synthases of the Shikimate Pathway Display Both Manganese‐ and Cobalt‐Dependent Activities |
| title_sort | arabidopsis 3 deoxy d arabino heptulosonate 7 phosphate dahp synthases of the shikimate pathway display both manganese and cobalt dependent activities |
| url | https://doi.org/10.1002/pld3.70037 |
| work_keys_str_mv | AT ryoyokoyama arabidopsis3deoxydarabinoheptulosonate7phosphatedahpsynthasesoftheshikimatepathwaydisplaybothmanganeseandcobaltdependentactivities AT hiroshiamaeda arabidopsis3deoxydarabinoheptulosonate7phosphatedahpsynthasesoftheshikimatepathwaydisplaybothmanganeseandcobaltdependentactivities |