Engineering the cyanobacterial ATP-driven BCT1 bicarbonate transporter for functional targeting to C3 plant chloroplasts
© The Author(s) 2024. Published by Oxford University Press on behalf of the Society for Experimental Biology.
| Publié dans: | Journal of experimental botany. - 1985. - 75(2024), 16 vom: 28. Aug., Seite 4926-4943 |
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| Auteur principal: | |
| Autres auteurs: | , , , , , , , , , , |
| Format: | Article en ligne |
| Langue: | English |
| Publié: |
2024
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| Accès à la collection: | Journal of experimental botany |
| Sujets: | Journal Article ABC transporter CO2-concentrating mechanism bicarbonate transport chloroplast engineering chloroplast envelope improving photosynthesis Bacterial Proteins Bicarbonates Anion Transport Proteins plus... |
| Résumé: | © The Author(s) 2024. Published by Oxford University Press on behalf of the Society for Experimental Biology. The ATP-driven bicarbonate transporter 1 (BCT1) from Synechococcus is a four-component complex in the cyanobacterial CO2-concentrating mechanism. BCT1 could enhance photosynthetic CO2 assimilation in plant chloroplasts. However, directing its subunits (CmpA, CmpB, CmpC, and CmpD) to three chloroplast sub-compartments is highly complex. Investigating BCT1 integration into Nicotiana benthamiana chloroplasts revealed promising targeting strategies using transit peptides from the intermembrane space protein Tic22 for correct CmpA targeting, while the transit peptide of the chloroplastic ABCD2 transporter effectively targeted CmpB to the inner envelope membrane. CmpC and CmpD were targeted to the stroma by RecA and recruited to the inner envelope membrane by CmpB. Despite successful targeting, expression of this complex in CO2-dependent Escherichia coli failed to demonstrate bicarbonate uptake. We then used rational design and directed evolution to generate new BCT1 forms that were constitutively active. Several mutants were recovered, including a CmpCD fusion. Selected mutants were further characterized and stably expressed in Arabidopsis thaliana, but the transformed plants did not have higher carbon assimilation rates or decreased CO2 compensation points in mature leaves. While further analysis is required, this directed evolution and heterologous testing approach presents potential for iterative modification and assessment of CO2-concentrating mechanism components to improve plant photosynthesis |
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| Description: | Date Completed 28.08.2024 Date Revised 09.09.2024 published: Print Citation Status MEDLINE |
| ISSN: | 1460-2431 |
| DOI: | 10.1093/jxb/erae234 |