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• W3102551569 abstract "Abstract H 2 O 2 has been recognized as an important signaling molecule in plants. We sought to establish a genetically encoded, fluorescent H 2 O 2 sensor that allows H 2 O 2 monitoring in all major subcompartments of a Chlamydomonas cell. To this end we engineered the hypersensitive H 2 O 2 sensor, roGFP2-Tsa2ΔC R , as a genetic part for the Chlamydomonas Modular Cloning toolbox. Using previously generated parts, together with new ones, we constructed modules and devices that target the sensor to the cytosol, nucleus, mitochondrial matrix, chloroplast stroma, thylakoid lumen, and ER. The sensor was functional in all compartments, except for the ER where it was fully oxidized. Employing our novel sensors, we show that H 2 O 2 produced by photosynthetic linear electron transport (PET) in the stroma leaks into the cytosol but only reaches other subcellular compartments if produced under non-physiological conditions. Our results thus imply the establishment of steep intracellular H 2 O 2 gradients under normal physiological conditions and suggest that the cytosolic complement of H 2 O 2 scavenging enzymes effectively limits H 2 O 2 diffusion. Furthermore, in heat stressed cells, we show that cytosolic H 2 O 2 levels closely mirror temperature up- and downshifts and are independent from PET. We anticipate that these sensors will greatly facilitate future investigations into H 2 O 2 biology in algal and plant cells." @default.
• W3102551569 created "2020-11-23" @default.
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• W3102551569 date "2020-11-16" @default.
• W3102551569 modified "2023-09-24" @default.
• W3102551569 title "Real-time monitoring of subcellular H₂O₂ distribution in <i>Chlamydomonas reinhardtii</i>" @default.
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• W3102551569 doi "https://doi.org/10.1101/2020.11.13.382085" @default.
• W3102551569 hasPublicationYear "2020" @default.
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