Conserved Chloroplast Open-reading Frame ycf54 Is Required for Activity of the Magnesium Protoporphyrin Monomethylester Oxidative Cyclase in Synechocystis PCC 6803

Hollingshead, Sarah; Kopečná, Jana; Jackson, Philip J.; Canniffe, Daniel P.; Davison, Paul A.; Dickman, Mark J.; Sobotka, Roman; Hunter, C. Neil

doi:10.1074/jbc.m112.352526

Cited by 81 publications

(116 citation statements)

References 33 publications

(41 reference statements)

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“…To generate the strains of Synechocystis sp PCC 6803 expressing YFP or a TorA-YFP fusion, genes encoding the respective proteins were cloned into the NdeI/BglII sites of the pPD-FLAG vector (Hollingshead et al, 2012), replacing the 3xFLAG tag coding sequence. When the resulting plasmids were introduced into Synechocystis (as described in Hollingshead et al, 2012), the inserted gene was integrated in place of psbAII (slr1311) by double homologous recombination.…”

Section: Growth Of Cells and Strain Generationmentioning

confidence: 99%

“…When the resulting plasmids were introduced into Synechocystis (as described in Hollingshead et al, 2012), the inserted gene was integrated in place of psbAII (slr1311) by double homologous recombination. Transformants were selected on BG-11 plates containing 5 mg/mL kanamycin, and genome copies were segregated by plating with sequential doubling of the kanamycin concentration up to 40 mg/mL.…”

Section: Growth Of Cells and Strain Generationmentioning

confidence: 99%

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Lateral Segregation of Photosystem I in Cyanobacterial Thylakoids

et al. 2017

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Photosystem I (PSI) is the dominant photosystem in cyanobacteria and it plays a pivotal role in cyanobacterial metabolism. Despite its biological importance, the native organization of PSI in cyanobacterial thylakoid membranes is poorly understood. Here, we use atomic force microscopy (AFM) to show that ordered, extensive macromolecular arrays of PSI complexes are present in thylakoids from Thermosynechococcus elongatus, Synechococcus sp PCC 7002, and Synechocystis sp PCC 6803. Hyperspectral confocal fluorescence microscopy and three-dimensional structured illumination microscopy of Synechocystis sp PCC 6803 cells visualize PSI domains within the context of the complete thylakoid system. Crystallographic and AFM data were used to build a structural model of a membrane landscape comprising 96 PSI trimers and 27,648 chlorophyll a molecules. Rather than facilitating intertrimer energy transfer, the close associations between PSI primarily maximize packing efficiency; short-range interactions with Complex I and cytochrome b 6 f are excluded from these regions of the membrane, so PSI turnover is sustained by long-distance diffusion of the electron donors at the membrane surface. Elsewhere, PSI-photosystem II contact zones provide sites for docking phycobilisomes and the formation of megacomplexes. PSI-enriched domains in cyanobacteria might foreshadow the partitioning of PSI into stromal lamellae in plants, similarly sustained by long-distance diffusion of electron carriers.

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Section: Growth Of Cells and Strain Generationmentioning

confidence: 99%

Section: Growth Of Cells and Strain Generationmentioning

confidence: 99%

Lateral Segregation of Photosystem I in Cyanobacterial Thylakoids

et al. 2017

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“…Gun4 was found to stimulate cyanobacterial MgCH activity in vitro (35) and to be involved in increasing flux into the Chl biosynthesis pathway in vivo (36). Similarly, it is conceivable that Ycf54 may play a role in substrate channeling; pull-down experiments identified protein-protein interactions between Ycf54 and the cyanobacterial AcsF, CycI (21), and CycI with other Chl biosynthesis enzymes (22). These interactions are abrogated in the absence of ycf54, and the level of CycI is reduced, suggesting that Ycf54 may stabilize the CycI protein (22).…”

Section: Discussionmentioning

confidence: 99%

“…Previous work has shown that Synechocystis Δycf54 makes a small amount of Chl, ∼13% of WT levels (21,22); thus, Ycf54 appears to be important, but not essential, for cyclase activity. To assess the contribution of Ycf54 more precisely, we developed a reciprocal system for the heterologous expression of Synechocystis genes in Rvi.…”

Section: Rvi Gelatinosus Acsf Complements the Loss Of Cyci In Synechmentioning

confidence: 90%

“…Later in vivo pulldown experiments in Synechocystis with tagged CycI and CycII identified Ycf54 as an interaction partner of both AcsF isoforms; tagged Ycf54 produced in the same manner was shown to interact with CycI (21). A mutant depleted in Ycf54 had a significantly reduced Chl content, accumulated MgPME, was only able to tolerate low light intensities and was unable to grow under photoautotrophic conditions (21,22). Antisense mutants of the ortholog of ycf54 in tobacco, LCAA, also demonstrated impaired cyclase activity, and the dimeric protein was shown to interact with CHL27 in the chloroplast (23).…”

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Three classes of oxygen-dependent cyclase involved in chlorophyll and bacteriochlorophyll biosynthesis

Chen

Canniffe

Hunter

2017

Proc. Natl. Acad. Sci. U.S.A.

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The biosynthesis of (bacterio)chlorophyll pigments is among the most productive biological pathways on Earth. Photosynthesis relies on these modified tetrapyrroles for the capture of solar radiation and its conversion to chemical energy. (Bacterio)chlorophylls have an isocyclic fifth ring, the formation of which has remained enigmatic for more than 60 y. This reaction is catalyzed by two unrelated cyclase enzymes using different chemistries. The majority of anoxygenic phototrophic bacteria use BchE, an O 2 -sensitive [4Fe-4S] cluster protein, whereas plants, cyanobacteria, and some phototrophic bacteria possess an O 2 -dependent enzyme, the major catalytic component of which is a diiron protein, AcsF. Plant and cyanobacterial mutants in ycf54 display impaired function of the O 2 -dependent enzyme, accumulating the reaction substrate. Swapping cyclases between cyanobacteria and purple phototrophic bacteria reveals three classes of the O 2 -dependent enzyme. AcsF from the purple betaproteobacterium Rubrivivax (Rvi.) gelatinosus rescues the loss not only of its cyanobacterial ortholog, cycI, in Synechocystis sp. PCC 6803, but also of ycf54; conversely, coexpression of cyanobacterial cycI and ycf54 is required to complement the loss of acsF in Rvi. gelatinosus. These results indicate that Ycf54 is a cyclase subunit in oxygenic phototrophs, and that different classes of the enzyme exist based on their requirement for an additional subunit. AcsF is the cyclase in Rvi. gelatinosus, whereas alphaproteobacterial cyclases require a newly discovered protein that we term BciE, encoded by a gene conserved in these organisms. These data delineate three classes of O 2 -dependent cyclase in chlorophototrophic organisms from higher plants to bacteria, and their evolution is discussed herein.

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Plant and algal chlorophyll synthases function in Synechocystis and interact with the YidC/Alb3 membrane insertase

et al. 2018

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In the model cyanobacterium Synechocystis sp. PCC 6803, the terminal enzyme of chlorophyll biosynthesis, chlorophyll synthase (ChlG), forms a complex with high light‐inducible proteins, the photosystem II assembly factor Ycf39 and the YidC/Alb3/OxaI membrane insertase, co‐ordinating chlorophyll delivery with cotranslational insertion of nascent photosystem polypeptides into the membrane. To gain insight into the ubiquity of this assembly complex in higher photosynthetic organisms, we produced functional foreign chlorophyll synthases in a cyanobacterial host. Synthesis of algal and plant chlorophyll synthases allowed deletion of the otherwise essential native cyanobacterial gene. Analysis of purified protein complexes shows that the interaction with YidC is maintained for both eukaryotic enzymes, indicating that a ChlG‐YidC/Alb3 complex may be evolutionarily conserved in algae and plants.

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Conserved Chloroplast Open-reading Frame ycf54 Is Required for Activity of the Magnesium Protoporphyrin Monomethylester Oxidative Cyclase in Synechocystis PCC 6803

Cited by 81 publications

References 33 publications

Lateral Segregation of Photosystem I in Cyanobacterial Thylakoids

Lateral Segregation of Photosystem I in Cyanobacterial Thylakoids

Three classes of oxygen-dependent cyclase involved in chlorophyll and bacteriochlorophyll biosynthesis

Plant and algal chlorophyll synthases function in Synechocystis and interact with the YidC/Alb3 membrane insertase

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