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Plant J. 2019 Jul;99(1):176-194. doi: 10.1111/tpj.14315. Epub 2019 May 13.

Diurnal changes in concerted plant protein phosphorylation and acetylation in Arabidopsis organs and seedlings.

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Institute of Molecular Plant Biology, Department of Biology, ETH Zurich, 8092, Zurich, Switzerland.
Department of Biological Sciences, University of Alberta, Edmonton, Alberta, Canada.
Functional Genomics Center, ETH Zurich, 8092, Zurich, Switzerland.
Advanced Plant Biotechnology Center, National Chung Hsing University, Taichung, 40227, Taiwan.


Protein phosphorylation and acetylation are the two most abundant post-translational modifications (PTMs) that regulate protein functions in eukaryotes. In plants, these PTMs have been investigated individually; however, their co-occurrence and dynamics on proteins is currently unknown. Using Arabidopsis thaliana, we quantified changes in protein phosphorylation, acetylation and protein abundance in leaf rosettes, roots, flowers, siliques and seedlings at the end of day (ED) and at the end of night (EN). This identified 2549 phosphorylated and 909 acetylated proteins, of which 1724 phosphorylated and 536 acetylated proteins were also quantified for changes in PTM abundance between ED and EN. Using a sequential dual-PTM workflow, we identified significant PTM changes and intersections in these organs and plant developmental stages. In particular, cellular process-, pathway- and protein-level analyses reveal that the phosphoproteome and acetylome predominantly intersect at the pathway- and cellular process-level at ED versus EN. We found 134 proteins involved in core plant cell processes, such as light harvesting and photosynthesis, translation, metabolism and cellular transport, that were both phosphorylated and acetylated. Our results establish connections between PTM motifs, PTM catalyzing enzymes and putative substrate networks. We also identified PTM motifs for further characterization of the regulatory mechanisms that control cellular processes during the diurnal cycle in different Arabidopsis organs and seedlings. The sequential dual-PTM analysis expands our understanding of diurnal plant cell regulation by PTMs and provides a useful resource for future analyses, while emphasizing the importance of analyzing multiple PTMs simultaneously to elucidate when, where and how they are involved in plant cell regulation.


Arabidopsis thaliana ; acetylation; diurnal cycle; phosphorylation; proteomics


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