Immobilization of a Broad Range of Polypeptides on the Frustule of the Diatom<i>Thalassiosira pseudonana</i>

Shimakawa, Ginga; Katayama, Shota; Tsuji, Yoshinori; Yoneda, Kohei; Fukuda, Wakao; Fujiwara, Shinsuke; Matsuda, Yusuke

doi:10.1128/aem.01153-22

Cited by 3 publications

(2 citation statements)

References 37 publications

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“…Chlorophyll autofluorescence was evaluated from the emission between 600 and 750 nm excited by a 552 nm laser. GFP was excited by a 488 nm laser, and the green fluorescence was detected at 500−520 nm (Shimakawa et al, 2022).…”

Section: Methodsmentioning

confidence: 99%

Low-CO₂inducible bestrophins in diatom thylakoid membranes sustain high photosynthetic efficacy at distant locations from the pyrenoid

Nigishi,

Shimakawa,

Yamagishi

et al. 2023

Preprint

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Anion transporters are important to sustain a variety of physiological states in cells. Bestrophins are a family of Cl- and/or HCO3- transporters conserved in bacteria, animals, algae, and plants. Recently, bestrophin paralogs were found in the green alga Chlamydomonas reinhardtii as up-regulated components in low CO2 conditions that play an essential role in the CO2-concentrating mechanism (CCM). Bestrophin orthologs are also conserved in diatoms, a group of secondary endosymbiotic algae harboring red-type plastids, but their physiological functions are not known yet. Here, we characterized the subcellular localization and expression profile of bestrophins in the marine diatoms Phaeodactylum tricornutum (PtBST1-4) and Thalassiosira pseudonana (TpBST1 and 2). PtBST1 and PtBST2 were localized at the stromal thylakoid membrane outside of the pyrenoid, and PtBST3 was localized in the pyrenoid. Contrarily, TpBST1 and TpBST2 were both localized in the pyrenoid. These bestrophin proteins were accumulated in cells grown in atmospheric CO2 but not in 1% CO2-grown cells. To assess the physiological functions, we generated knock-out mutants for PtBST1 by genome editing. The lack of PtBST1 decreased affinity of photosynthesis for dissolved inorganic carbon closer to that of the cells grown in 1% CO2. Additionally, non-photochemical quenching was 1.5-2.0 times higher in the mutants than that of the wild type cells. These data suggests that HCO3- transport at the stroma thylakoid membranes by PtBST1 is a critical part of the CO2 evolving system of the pyrenoid in the fully induced CCM, and simultaneously that PtBST1 modulates photoprotection in response to CO2 availability in P. tricornutum.

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Section: Methodsmentioning

confidence: 99%

Low-CO₂inducible bestrophins in diatom thylakoid membranes sustain high photosynthetic efficacy at distant locations from the pyrenoid

Nigishi,

Shimakawa,

Yamagishi

et al. 2023

Preprint

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“…When the recombinant genes were expressed, the resulting fusion proteins became stably immobilized into the T. pseudonana cell wall, and the enzyme or receptor domains retained their functions. The silica‐immobilized enzymes had enhanced stability against denaturing conditions and have potential as reusable catalysts in the pharmaceutical industry, food processing, cancer treatment, chemical sensing, or wastewater treatment (Delalat et al, 2015; Kröger et al, 2017; Kumari et al, 2020; Marshall et al, 2012; Poulsen et al, 2007; Shimakawa et al, 2022; Xiong et al, 2016).…”

Section: Silica Biotechnologymentioning

confidence: 99%

Thalassiosira pseudonana (Cyclotella nana) (Hustedt) Hasle et Heimdal (Bacillariophyceae): A genetically tractable model organism for studying diatom biology, including biological silica formation

Poulsen

Kröger

2023

Journal of Phycology

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In 2004, Thalassiosira pseudonana was the first eukaryotic marine alga to have its genome sequenced. Since then, this species has quickly emerged as a valuable model species for investigating the molecular underpinnings of essentially all aspects of diatom life, particularly bio‐morphogenesis of the cell wall. An important prerequisite for the model status of T. pseudonana is the ongoing development of increasingly precise tools to study the function of gene networks and their encoded proteins in vivo. Here, we briefly review the current toolbox for genetic manipulation, highlight specific examples of its application in studying diatom metabolism, and provide a peek into the role of diatoms in the emerging field of silica biotechnology.

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Low-CO2-inducible bestrophins outside the pyrenoid sustain high photosynthetic efficacy in diatoms

Nigishi,

Shimakawa,

Yamagishi

et al. 2024

Plant Physiology

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Anion transporters sustain a variety of physiological states in cells. Bestrophins belong to a Cl− and/or HCO3− transporter family conserved in bacteria, animals, algae, and plants. Recently, putative bestrophins were found in the green alga Chlamydomonas reinhardtii, where they are up-regulated under low CO2 conditions and play an essential role in the CO2-concentrating mechanism (CCM). The putative bestrophin orthologs are also conserved in diatoms, secondary endosymbiotic algae harboring red-type plastids, but their physiological functions are unknown. Here, we characterized the subcellular localization and expression profile of bestrophins (BSTs) in the marine diatoms Phaeodactylum tricornutum (PtBST1−4) and Thalassiosira pseudonana (TpBST1 and 2). PtBST1, PtBST2, and PtBST4 localized at the stroma thylakoid membrane outside of the pyrenoid, and PtBST3 localized in the pyrenoid. Contrarily, TpBST1 and TpBST2 both localized in the pyrenoid. These bestrophin proteins accumulated in cells grown in atmospheric CO2 (LC) but not in 1% CO2 (HC)-grown cells. To assess the physiological functions, we generated knock-out mutants for the PtBST1gene by genome editing. The lack of PtBST1 decreased photosynthetic affinity for dissolved inorganic carbon to the level comparable to the HC-grown wild type. Furthermore, non-photochemical quenching in LC-grown cells was 1.5–2.0 times higher in the mutants than in the wild type. These data suggest that HCO3− transport at the stroma thylakoid membranes by PtBST1 is a critical part of the CO2-evolving machinery of the pyrenoid in the fully induced CCM and that PtBST1 may modulate photoprotection under CO2-limited environments in P. tricornutum.

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Immobilization of a Broad Range of Polypeptides on the Frustule of the DiatomThalassiosira pseudonana

Cited by 3 publications

References 37 publications

Low-CO₂inducible bestrophins in diatom thylakoid membranes sustain high photosynthetic efficacy at distant locations from the pyrenoid

Low-CO₂inducible bestrophins in diatom thylakoid membranes sustain high photosynthetic efficacy at distant locations from the pyrenoid

Thalassiosira pseudonana (Cyclotella nana) (Hustedt) Hasle et Heimdal (Bacillariophyceae): A genetically tractable model organism for studying diatom biology, including biological silica formation

Low-CO2-inducible bestrophins outside the pyrenoid sustain high photosynthetic efficacy in diatoms

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Immobilization of a Broad Range of Polypeptides on the Frustule of the DiatomThalassiosira pseudonana

Cited by 3 publications

References 37 publications

Low-CO2inducible bestrophins in diatom thylakoid membranes sustain high photosynthetic efficacy at distant locations from the pyrenoid

Low-CO2inducible bestrophins in diatom thylakoid membranes sustain high photosynthetic efficacy at distant locations from the pyrenoid

Thalassiosira pseudonana (Cyclotella nana) (Hustedt) Hasle et Heimdal (Bacillariophyceae): A genetically tractable model organism for studying diatom biology, including biological silica formation

Low-CO2-inducible bestrophins outside the pyrenoid sustain high photosynthetic efficacy in diatoms

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Low-CO₂inducible bestrophins in diatom thylakoid membranes sustain high photosynthetic efficacy at distant locations from the pyrenoid

Low-CO₂inducible bestrophins in diatom thylakoid membranes sustain high photosynthetic efficacy at distant locations from the pyrenoid