Chen, G.E., Hitchcock, A. orcid.org/0000-0001-6572-434X, Mareš, J. et al. (8 more authors) (2021) Evolution of Ycf54-independent chlorophyll biosynthesis in cyanobacteria. Proceedings of the National Academy of Sciences, 118 (10). e2024633118. ISSN 0027-8424
Abstract
Chlorophylls (Chls) are essential cofactors for photosynthesis. One of the least understood steps of Chl biosynthesis is formation of the fifth (E) ring, where the red substrate, magnesium protoporphyrin IX monomethyl ester, is converted to the green product, 3,8-divinyl protochlorophyllide a. In oxygenic phototrophs, this reaction is catalyzed by an oxygen-dependent cyclase, consisting of a catalytic subunit (AcsF/CycI) and an auxiliary protein, Ycf54. Deletion of Ycf54 impairs cyclase activity and results in severe Chl deficiency, but its exact role is not clear. Here, we used a Δycf54 mutant of the model cyanobacterium Synechocystis sp. PCC 6803 to generate suppressor mutations that restore normal levels of Chl. Sequencing Δycf54 revertants identified a single D219G amino acid substitution in CycI and frameshifts in slr1916, which encodes a putative esterase. Introduction of these mutations to the original Δycf54 mutant validated the suppressor effect, especially in combination. However, comprehensive analysis of the Δycf54 suppressor strains revealed that the D219G-substituted CycI is only partially active and its accumulation is misregulated, suggesting that Ycf54 controls both the level and activity of CycI. We also show that Slr1916 has Chl dephytylase activity in vitro and its inactivation up-regulates the entire Chl biosynthetic pathway, resulting in improved cyclase activity. Finally, large-scale bioinformatic analysis indicates that our laboratory evolution of Ycf54-independent CycI mimics natural evolution of AcsF in low-light–adapted ecotypes of the oceanic cyanobacteria Prochlorococcus, which lack Ycf54, providing insight into the evolutionary history of the cyclase enzyme.
Metadata
Item Type: | Article |
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Authors/Creators: |
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Copyright, Publisher and Additional Information: | © 2021 Published under the PNAS license. This is an author-produced version of a paper subsequently published in Proceedings of the National Academy of Sciences of the USA. Uploaded in accordance with the publisher's self-archiving policy. |
Keywords: | photosynthesis; chlorophyll; cyclase; cyanobacteria; microevolution |
Dates: |
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Institution: | The University of Sheffield |
Academic Units: | The University of Sheffield > Faculty of Science (Sheffield) > School of Biosciences (Sheffield) |
Funding Information: | Funder Grant number BIOTECHNOLOGY AND BIOLOGICAL SCIENCES RESEARCH COUNCIL BB/M000265/1 EUROPEAN COMMISSION - HORIZON 2020 854126 |
Depositing User: | Symplectic Sheffield |
Date Deposited: | 28 Oct 2021 08:40 |
Last Modified: | 10 Feb 2023 14:11 |
Status: | Published |
Publisher: | Proceedings of the National Academy of Sciences |
Refereed: | Yes |
Identification Number: | 10.1073/pnas.2024633118 |
Related URLs: | |
Open Archives Initiative ID (OAI ID): | oai:eprints.whiterose.ac.uk:179709 |