Dataset Information

PSI supercomplex copurification using PSI His-tag

ABSTRACT: Chlamydomonas reinhardtii PSI-Cyt b6f complexes from sucrose density gradient fractions were identified by western blot and SMF spectroscopy, but showed high heterogeneity, as evident from the multiple bands present in the Coomassie stained SDS gel and original EM micrographs. Therefore potential PSI-Cyt b6f supercomplexes were enriched by a subsequent PsaA HisTag purification step. Only this additional enrichment enabled the identification of novel PSI supercomplexes, although the sample remained far from being homogeneous. This dataset describes the quantitative mass spectrometric analysis of the enriched PSI supercomplex fraction in comparison to unpurified samples using metabolic 15N labeling.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Chlamydomonas Reinhardtii

TISSUE(S): Photosynthetic Cell

SUBMITTER: Stefan Schulze

LAB HEAD: Michael Hippler

PROVIDER: PXD008857 | Pride | 2018-10-03

REPOSITORIES: Pride

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Publications

Structure of a PSI-LHCI-cyt b<sub>6</sub>f supercomplex in <i>Chlamydomonas reinhardtii</i> promoting cyclic electron flow under anaerobic conditions.

Steinbeck Janina J Ross Ian L IL Rothnagel Rosalba R Gäbelein Philipp P Schulze Stefan S Giles Nichole N Ali Rubbiya R Drysdale Rohan R Sierecki Emma E Gambin Yann Y Stahlberg Henning H Takahashi Yuichiro Y Hippler Michael M Hankamer Ben B

Proceedings of the National Academy of Sciences of the United States of America 20180925 41

Photosynthetic linear electron flow (LEF) produces ATP and NADPH, while cyclic electron flow (CEF) exclusively drives photophosphorylation to supply extra ATP. The fine-tuning of linear and cyclic electron transport levels allows photosynthetic organisms to balance light energy absorption with cellular energy requirements under constantly changing light conditions. As LEF and CEF share many electron transfer components, a key question is how the same individual structural units contribute to the ...[more]

PMID: 30254175

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Project description:"Background: The ascomycete Fusarium fujikuroi causes “bakanae” disease of rice due to its ability to produce gibberellins (GA), but is also known for producing a broad spectrum of other secondary metabolites, (SM) such as pigments and harmful mycotoxins. Biosynthesis of multiple SM is regulated by nitrogen availability. The global GATA transcription factors (TF) AreA and AreB were shown to play key roles in this nitrogen-dependent regulation. Recently, we have shown that AreA activates the transcription of AREB, but not vice versa. Both regulators interact in the nucleus, indicating a complex regulatory interplay between both regulators. Results: We performed transcriptome and proteome analyses of the wild type and ΔAREA and ΔAREB mutants under nitrogen limiting and sufficient conditions. Microarray analysis revealed that each transcription factor regulates a total set of about 4,200 genes under nitrogen limiting conditions. Gene enrichment analysis revealed that significantly up- and down-regulated genes belong to functional classes of primary/secondary metabolism, transporters, cell defense/virulence and transcription. Under nitrogen excess, AreA is not active as a transcriptional regulator, while AreB affects expression of about 4,400 genes. Both transcription factors regulate a common set of about 2,100 genes, often coordinately either as positive or as negative regulators. Our proteomics analysis identified 446 AreA-regulated and 386 AreB-regulated proteins under nitrogen limitation and 138 AreA-regulated and 447 AreB-regulated proteins under nitrogen sufficiency, respectively. Comparison of the proteome and transcriptome data indicates that both GATA-factors not only regulate expression of target genes but also affect protein abundance by post-transcriptional regulations, for example on enzymes involved in glutamate cycling, as well as histone acetylation at some of the target SM gene clusters. Conclusion: This study provides novel insights into the AreA- and AreB-mediated regulation network on a cellular scale. Microarray analysis clearly demonstrated that both GATA-factors are major regulators of alternate nitrogen assimilatory pathways and secondary metabolism. Moreover, proteome analysis of the AREA and AREB deletion mutants showed that both TF are global players interconnecting carbon and nitrogen regulation. They mainly act as mutualistic, but also antagonistic regulators and operate by transcriptional and post-transcriptional regulations and epigenetic modifications." (Abstract taken from manuscript)

Dataset Information

PSI supercomplex copurification using PSI His-tag

Publications

Structure of a PSI-LHCI-cyt b<sub>6</sub>f supercomplex in <i>Chlamydomonas reinhardtii</i> promoting cyclic electron flow under anaerobic conditions.

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