Dataset Information

Phototropin-Mediated Signaling Kinase regulation in Chlamydomonas reinhardtii

ABSTRACT: Green algae, such as Chlamydomonas reinhardtii, use light as an energy source, as well as a source of information. Light is processed by photoreceptors, such as phototropin, which subsequently send signals throughout the organism. Herein, a phosphoproteomic workflow is employed to charachterize signaling pathways of phototropin. For phosphopeptide enrichment, tryptic digests were diluted with 100 µL of acetonitrile. A Fe-IMAC column was equilibrated, and each sample was injected separately onto the column.

INSTRUMENT(S):

ORGANISM(S): Chlamydomonas Reinhardtii

SUBMITTER: Leslie Hicks

LAB HEAD: Leslie Hicks

PROVIDER: PXD045599 | Pride | 2025-02-03

REPOSITORIES: Pride

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			Action	DRS
	20230508_PWS_DIMITRIS_CR_WT_B_4.timeseries.data	Other
	20230508_PWS_DIMITRIS_CR_WT_B_4.wiff	Wiff
	20230508_PWS_DIMITRIS_CR_WT_B_4.wiff.scan	Wiff
	20230508_PWS_DIMITRIS_CR_WT_B_4.wiff2	Wiff
	20230512_PWS_DIMITRIS_CR_PHOT_B_1.timeseries.data	Other

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Project description:Pelagic aggregates function as hotspots for microbial activity and biological carbon pumps for exporting OM fixed by photoautotrophs to sediments in lakes and oceans. In iron-rich (ferruginous) lakes, photoferrotrophic or chemolithoautotrophic bacteria appear to contribute to CO2 fixation by oxidizing reduced iron which leads to the formation of iron-rich pelagic aggregates called iron-snow. In acidic lakes, iron snow is colonized mainly by acidophilic iron-cycling microbes that can trigger interspecies aggregation mechanisms. However, the significance of iron oxidizers in carbon fixation, their general role in iron snow functioning, and the flow of carbon within iron snow is still unclear. Here, we combined a two-year metatranscriptome analysis with a 13CO2 metabolic labeling approach to determine general metabolic activities. Protein-based stable isotope probing (protein-SIP) was used to trace the 13CO2 incorporation in iron snow microcosms over time under both oxic and anoxic conditions. Analysis of our mRNA-derived metatranscriptome data identified four key players (Leptospirillum, Ferrovum, Acidithrix, Acidiphilium) with relative abundances (59.6%-85.7%) in iron snow encoding a variety of ecologically relevant pathways, including carbon fixation, polysaccharide biosynthesis, and flagellar-based motility. We did not detect transcriptional activity for carbon fixation from archaea or eukaryotes. The largest numbers of expressed genes (3008, 2991, 2936) matched to the genomes of our previously obtained iron snow isolates (Acidithrix sp. C25, Acidiphilium sp. C61, Acidocella sp. C78) separately. 13CO2 incorporation studies identified Leptospirillum and Ferrovum, as the main active chemolithoautotrophs under oxic conditions, and Ferrovum was the main active organism under anoxic conditions as well. Small amounts of labeled 13C (Relative isotope abundance: 1.0%-5.3%) were found in the heterotrophic Acidiphilium and Acidocella. Overall, our data show that iron oxidizers play an important role in the formation of iron minerals and CO2 fixation, but the majority of fixed C apparently did not reach other iron snow microbes. This finding suggests that most of the fixed C will be directly exported to the sediment without feeding heterotrophs in the water column in acidic ferruginous lakes.

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Phototropin-Mediated Signaling Kinase regulation in Chlamydomonas reinhardtii

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