Journal: Cell / Year: 2024 Title: Diatom pyrenoids are encased in a protein shell that enables efficient CO fixation. Authors: Ginga Shimakawa / Manon Demulder / Serena Flori / Akihiro Kawamoto / Yoshinori Tsuji / Hermanus Nawaly / Atsuko Tanaka / Rei Tohda / Tadayoshi Ota / Hiroaki Matsui / Natsumi Morishima / ...Authors: Ginga Shimakawa / Manon Demulder / Serena Flori / Akihiro Kawamoto / Yoshinori Tsuji / Hermanus Nawaly / Atsuko Tanaka / Rei Tohda / Tadayoshi Ota / Hiroaki Matsui / Natsumi Morishima / Ryosuke Okubo / Wojciech Wietrzynski / Lorenz Lamm / Ricardo D Righetto / Clarisse Uwizeye / Benoit Gallet / Pierre-Henri Jouneau / Christoph Gerle / Genji Kurisu / Giovanni Finazzi / Benjamin D Engel / Yusuke Matsuda / Abstract: Pyrenoids are subcompartments of algal chloroplasts that increase the efficiency of Rubisco-driven CO fixation. Diatoms fix up to 20% of global CO, but their pyrenoids remain poorly characterized. ...Pyrenoids are subcompartments of algal chloroplasts that increase the efficiency of Rubisco-driven CO fixation. Diatoms fix up to 20% of global CO, but their pyrenoids remain poorly characterized. Here, we used in vivo photo-crosslinking to identify pyrenoid shell (PyShell) proteins, which we localized to the pyrenoid periphery of model pennate and centric diatoms, Phaeodactylum tricornutum and Thalassiosira pseudonana. In situ cryo-electron tomography revealed that pyrenoids of both diatom species are encased in a lattice-like protein sheath. Single-particle cryo-EM yielded a 2.4-Å-resolution structure of an in vitro TpPyShell1 lattice, which showed how protein subunits interlock. T. pseudonana TpPyShell1/2 knockout mutants had no PyShell sheath, altered pyrenoid morphology, and a high-CO requiring phenotype, with reduced photosynthetic efficiency and impaired growth under standard atmospheric conditions. The structure and function of the diatom PyShell provide a molecular view of how CO is assimilated in the ocean, a critical ecosystem undergoing rapid change.
Name: T. pseudonana / type: cell / ID: 1 / Parent: 0 / Details: Pyrenoid in T. pseudonana chloroplast
Source (natural)
Organism: Thalassiosira pseudonana (Diatom)
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Experimental details
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Structure determination
Method
cryo EM
Processing
electron tomography
Aggregation state
cell
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Sample preparation
Buffer
pH: 7
Vitrification
Cryogen name: ETHANE
Sectioning
Focused ion beam - Instrument: OTHER / Focused ion beam - Ion: OTHER / Focused ion beam - Voltage: 3 / Focused ion beam - Current: 0.03 / Focused ion beam - Duration: 600 / Focused ion beam - Temperature: 83 K / Focused ion beam - Initial thickness: 1000 / Focused ion beam - Final thickness: 150 Focused ion beam - Details: The value given for _em_focused_ion_beam.instrument is FEI Aquilos II. This is not in a list of allowed values {'OTHER', 'DB235'} so OTHER is written into the XML file.
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Electron microscopy
Microscope
FEI TITAN KRIOS
Image recording
Film or detector model: FEI FALCON II (4k x 4k) / Detector mode: COUNTING / Average electron dose: 2.0 e/Å2
Electron beam
Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
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