Dataset Information

Plastoglobules BN-PAGE experiment (Arabidopsis PGs under unstress and high light conditions)

ABSTRACT: Proteome comparison of isolated plastoglobules (PGs) from Arabidopsis thaliana (Col-0 ecotype) under unstress (UNS) and high light (HL) stress conditions. For both conditions, PGs were isolated from three independent set of plants each (i.e. three for UNS and three for HL). PGs were isolated from the plants when they were 4 weeks old under long day photoperiod (16 hours of light and 8 darkness). The light stress was applied to the designated set of plants 5 days prior the completion of the 4 weeks by shifting the light intensity from 120 uE to 500 E. The PGs were solubilized with 1.1 % n-Dodecyl--D-Maltoside and separated in a Blue-Native gel. After the electrophoretic separation, gel lanes were cut and further divided into 10 independent slices corresponding to different molecular weights. These gel slices were subjeted to in-gel digestion with trypsin and the resulting peptides labeled with TMT reagents. Labeled peptides were mixed 1:1 and purified by solid phase extraction using c18 StageTips. Eluted peptides were sprayed into a ThermoScientific Q-Exactive HF-X mass spectrometer. Survey scans were taken in the Orbi trap (120000 resolution, determined at m/z 200) and the top 20 ions in each survey scan are then subjected to automatic higher energy collision induced dissociation (HCD) with fragment spectra acquired at 45000 resolution. Additionally, PG samples that were not separated in a Blue-native gel were trypsin digested and subjected for mass-spectrometry analysis as previously described.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Arabidopsis Thaliana (mouse-ear Cress)

TISSUE(S): Photosynthetic Cell, Leaf, Rosette

SUBMITTER: Roberto Espinoza Corral

LAB HEAD: Peter K. Lundquist

PROVIDER: PXD019735 | Pride | 2021-04-13

REPOSITORIES: Pride

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Publications

Molecular changes of Arabidopsis thaliana plastoglobules facilitate thylakoid membrane remodeling under high light stress.

Espinoza-Corral Roberto R Schwenkert Serena S Lundquist Peter K PK

The Plant journal : for cell and molecular biology 20210503 6

Plants require rapid responses to adapt to environmental stresses. This includes dramatic changes in the size and number of plastoglobule lipid droplets within chloroplasts. Although the morphological changes of plastoglobules are well documented, little is known about the corresponding molecular changes. To address this gap, we have compared the quantitative proteome, oligomeric state, prenyl-lipid content and kinase activities of Arabidopsis thaliana plastoglobules under unstressed and 5-day l ...[more]

PMID: 33783866

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Project description:Plastids contain multiple copies of the plastid chromosome, folded together with proteins and RNA into nucleoids. The degree to which components of the plastid gene expression and protein biogenesis machineries are nucleoid associated, and the factors involved in plastid DNA organization, repair, and replication, are poorly understood. To provide a conceptual framework for nucleoid function, we characterized the proteomes of highly enriched nucleoid fractions of proplastids and mature chloroplasts isolated from the maize (Zea mays) leaf base and tip, respectively, using mass spectrometry. Quantitative comparisons with proteomes of unfractionated proplastids and chloroplasts facilitated the determination of nucleoid-enriched proteins. This nucleoid-enriched proteome included proteins involved in DNA replication, organization, and repair as well as transcription, mRNA processing, splicing, and editing. Many proteins of unknown function, including pentatricopeptide repeat (PPR), tetratricopeptide repeat (TPR), DnaJ, and mitochondrial transcription factor (mTERF) domain proteins, were identified. Strikingly, 70S ribosome and ribosome assembly factors were strongly overrepresented in nucleoid fractions, but protein chaperones were not. Our analysis strongly suggests that mRNA processing, splicing, and editing, as well as ribosome assembly, take place in association with the nucleoid, suggesting that these processes occur cotranscriptionally. The plastid developmental state did not dramatically change the nucleoid-enriched proteome but did quantitatively shift the predominating function from RNA metabolism in undeveloped plastids to translation and homeostasis in chloroplasts. This study extends the known maize plastid proteome by hundreds of proteins, including more than 40 PPR and mTERF domain proteins, and provides a resource for targeted studies on plastid gene expression. Details of protein identification and annotation are provided in the Plant Proteome Database.

Dataset Information

Plastoglobules BN-PAGE experiment (Arabidopsis PGs under unstress and high light conditions)

Publications

Molecular changes of Arabidopsis thaliana plastoglobules facilitate thylakoid membrane remodeling under high light stress.

Similar Datasets

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