Nucleolar Proteome Analysis and Proteasomal Activity Assays Reveal a Link between Nucleolus and 26S Proteasome in A. thaliana

Frontiers in Plant Science
Charlotte MontaciéJulio Sáez-Vásquez

Abstract

In all eukaryotic cells, the nucleolus is functionally and structurally linked to rRNA synthesis and ribosome biogenesis. This compartment contains as well factors involved in other cellular activities, but the functional interconnection between non-ribosomal activities and the nucleolus (structure and function) still remains an open question. Here, we report a novel mass spectrometry analysis of isolated nucleoli from Arabidopsis thaliana plants using the FANoS (Fluorescence Assisted Nucleolus Sorting) strategy. We identified many ribosome biogenesis factors (RBF) and proteins non-related with ribosome biogenesis, in agreement with the recognized multi-functionality of the nucleolus. Interestingly, we found that 26S proteasome subunits localize in the nucleolus and demonstrated that proteasome activity and nucleolus organization are intimately linked to each other. Proteasome subunits form discrete foci in the disorganized nucleolus of nuc1.2 plants. Nuc1.2 protein extracts display reduced proteasome activity in vitro compared to WT protein extracts. Remarkably, proteasome activity in nuc1.2 is similar to proteasome activity in WT plants treated with proteasome inhibitors (MG132 or ALLN). Finally, we show that MG132 treatment ...Continue Reading

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Citations

Mar 15, 2019·Nucleus·Ariadna Picart-PicoloFrédéric Pontvianne
Apr 3, 2020·Journal of Plant Research·Frédéric Pontvianne, Stefan Grob
Jun 20, 2020·Journal of Experimental Botany·Ana Paula SantosSara Farrona
Jun 13, 2018·Frontiers in Plant Science·Laura MartinsJean-Philippe Reichheld
Dec 5, 2018·Chromosome Research : an International Journal on the Molecular, Supramolecular and Evolutionary Aspects of Chromosome Biology·Farah Bughio, Keith A Maggert
Aug 1, 2018·Nature Plants·Ezgi Süheyla Doğan, Chang Liu
Dec 15, 2020·The Plant Journal : for Cell and Molecular Biology·Konstantin O KutashevMartina Dvořáčková
Apr 2, 2021·Plant Physiology·Pei-Jung HsuWan-Hsing Cheng

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Methods Mentioned

BETA
Fluorescence
deubiquitination
FACS
Assay

Software Mentioned

msconvert
Proline
Mascot
FANoS
XCalibur

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