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Posewitz, M.

Publications and source records attributed to Posewitz, M..

3 recordsLinked to original sources

Enzyme-Constrained Metabolic Modeling Reveals CO2-Dependent Growth Control in Picochlorum celeri

Picochlorum celeri grows 11.6 times faster at elevated CO2 than in air, yet the abundance of its metabolic enzymes is only 17% higher. To explain this, we reconstructed iPiccel954, a genome-scale metabolic model with 3525 reactions, 2973 metabolites and 954 genes, and eciPiccel954, its enzyme-constrained version limited by the measured enzyme abundance in air and at elevated CO2. Calibrating the enzyme-constrained models against the measured proteome showed that a similar protein budget supports very different growth depending on how it is allocated. With urea as the nitrogen source, urea carboxylase was the only step whose relaxation alone raised growth, by 627% in air and 78% at elevated CO2. Bottleneck analysis showed that growth is limited by many enzymes spread across the network in both conditions, that the limiting enzymes differ between the two, and that control over starch metabolism seen in air disappears at elevated CO2. Measured proteomes agreed with these predictions, since the predicted bottleneck enzymes were more abundant than other enzymes. Rapid growth in P. celeri therefore reflects how the protein budget is allocated more than how large it is. Therefore, eciPiccel954 provides a quantitative basis for engineering this alga for biotechnological application.

systems biology↗

Multiomics dissection of the CO2-dependent fast growth of Picochlorum celeri

With a sub-3-micrometer cell and a 27-Mbp diploid genome, the small but mighty Picochlorum celeri doubles within three hours and exceeds 30 g m-2 d-1 outdoors with CO2 supplied. In climate-simulating photobioreactors, productivity declined from 39 g m-2 d-1 under 2.25% CO2 to near zero in air and resumed within hours of CO2 restoration with photosystem II photochemistry largely retained. We profiled the transcriptome, proteome, phosphoproteome, ubiquitinome, acetylome, metabolome and lipidome, including the first site-resolved green-algal ubiquitinome. In air, the Rubisco large subunit tripled to 7.6% of protein while its two small-subunit isoforms, 70% identical, exchanged near-reciprocally, a novel CO2-dependent switch that we propose retunes Rubisco to low CO2. Photorespiratory enzymes increased in abundance whereas proteins involved in external nitrogen assimilation decreased, consistent with increased reliance on internal nitrogen recycling as carbon fixation became limiting. Transcript and protein responses were nearly uncoupled, with cytosolic ribosomal proteins declining despite increased transcripts and plastid ribosomal proteins spared. Phosphorylation was extensively remodeled, as were the lysine modifications ubiquitination, acetylation and CO2-dependent carbamylation, where a CO2-competition assay identified 86 bicarbonate-protected lysines, including RbcL K252. CO2 limitation therefore imposes a reversible growth arrest that reduces cytosolic translational investment while preserving photosynthetic capacity for rapid recovery.

plant biology↗

Chlamydomonas reinhardtii triose-phosphate/phosphate translocator3 (TPT3): A major chloroplast shunt for the export of fixed carbon and reductant

Modulation of export of photoassimilates from the chloroplast is essential for controlling the distribution of fixed carbon in the cell and maintaining optimum photosynthetic rates. In this study we identified chloroplast triose phosphate/phosphate translocators 2 and 3 (CreTPT2 and CreTPT3) in the green alga Chlamydomonas reinhardtii that exhibited similar substrate specificities but were differentially expressed over the diel cycle. We focused mostly on analyzing CreTPT3 because of its high level of expression and the severe phenotype exhibited by tpt3 relative to the tpt2 mutants. Null mutants for CreTPT3 had a pleiotropic phenotype that impacted growth, photosynthetic activities, metabolite profiles, carbon partitioning, and organelle-specific accumulation of H2O2. These analyses demonstrated that CreTPT3 is a dominant conduit on the chloroplast envelope for the transport of photoassimilate. In addition, CreTPT3 can serve as a safety valve that moves excess reductant out of the chloroplast and appears to be essential for preventing the cells from experiencing oxidative stress and accumulating of reactive oxygen species, even under low/moderate light intensities. Finally, our studies indicate subfunctionalization of the CreTPT transporters and suggest that there are differences in managing the export of photoassimilates from the chloroplasts of Chlamydomonas and vascular plants.

plant biology↗