Quantitative phosphoproteomic and system-level analysis of TOR inhibition unravel distinct organellar acclimation in chlamydomonas reinhardtii

Author(s)
Valentin Roustan, Wolfram Weckwerth
Abstract

Rapamycin is an inhibitor of the evolutionary conserved Target of Rapamycin (TOR) kinase which promotes and coordinates translation with cell growth and division. In heterotrophic organisms, TOR regulation is based on intra- and extracellular stimuli such as amino acids level and insulin perception. However, how plant TOR pathways have evolved to integrate plastid endosymbiosis is a remaining question. Despite the close association of the TOR signaling with the coordination between protein turn-over and growth, proteome and phosphoproteome acclimation to a rapamycin treatment have not yet been thoroughly investigated in Chlamydomonas reinhardtii. In this study, we have used in vivo label-free phospho-proteomic analysis to profile both protein and phosphorylation changes at 0, 24, and 48 h in Chlamydomonas cells treated with rapamycin. Using multivariate statistics we highlight the impact of TOR inhibition on both the proteome and the phosphoproteome. Two-way ANOVA distinguished differential levels of proteins and phosphoproteins in response either to culture duration and rapamycin treatment or combined effects. Finally, protein–protein interaction networks and functional enrichment analysis underlined the relation between plastid and mitochondrial metabolism. Prominent changes of proteins involved in sulfur, cysteine, and methionine as well as nucleotide metabolism on the one hand, and changes in the TCA cycle on the other highlight the interplay of chloroplast and mitochondria metabolism. Furthermore, TOR inhibition revealed changes in the endomembrane trafficking system. Phosphoproteomics data, on the other hand, highlighted specific differentially regulated phosphorylation sites for calcium-regulated protein kinases as well as ATG7, S6K, and PP2C. To conclude we provide a first combined Chlamydomonas proteomics and phosphoproteomics dataset in response to TOR inhibition, which will support further investigations.

Organisation(s)
Research Platform Vienna Metabolomics Center, Large-Instrument Facility for Mass Spectrometry in Life Sciences
Journal
Frontiers in Plant Science
Volume
9
ISSN
1664-462X
DOI
https://doi.org/10.3389/fpls.2018.01590
Publication date
01-2018
Peer reviewed
Yes
Austrian Fields of Science 2012
106037 Proteomics, 106044 Systems biology, 106002 Biochemistry
Keywords
ASJC Scopus subject areas
Control and Systems Engineering, Computer Science(all)
Portal url
https://ucris.univie.ac.at/portal/en/publications/quantitative-phosphoproteomic-and-systemlevel-analysis-of-tor-inhibition-unravel-distinct-organellar-acclimation-in-chlamydomonas-reinhardtii(246f2189-cbdb-4dd6-90bd-6ea9b41a465c).html