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Abstract
Cytosolic 80S ribosomes contain proteins of the mature cytosolic ribosome (r-proteins) as well as proteins with roles in ribosome biogenesis, protein folding or modification. Here we refined the core r-protein composition in Arabidopsis thaliana by determining the abundance of different proteins during enrichment of ribosomes from cell cultures using peptide mass spectrometry. The turnover rates of 26 40S subunit r-proteins and 29 60S subunit r-proteins were also determined, showing that half of the ribosome population is replaced every 3-4 days. Three enriched proteins showed significantly shorter half-lives; a protein annotated as a ribosomal protein uL10 (RPP0D, At1g25260) with a half-life of 0.5 days and RACK1b and c with half-lives of 1-1.4 days. The At1g25260 protein is a homolog of the human Mrt4 protein, a trans-acting factor in the assembly of the pre-60S particle, while RACK1 has known regulatory roles in cell function beyond its role in the 40S subunit. Our experiments also identified 58 proteins that are not from r-protein families but co-purify with ribosomes and co-express with r-proteins; 26 were enriched more than 10-fold during ribosome enrichment. Some of these enriched proteins have known roles in translation, while others are newly proposed ribosome-associated factors in plants. This analysis provides an improved understanding of Arabidopsis thaliana ribosome protein content, shows that most r-proteins turnover in unison in vivo, identifies a novel set of potential plant translatome components, and how protein turnover can help identify r-proteins involved in ribosome biogenesis or regulation in plants.
Original language | English |
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Pages (from-to) | 3019-3032 |
Number of pages | 14 |
Journal | The Biochemical journal |
Volume | 477 |
Issue number | 16 |
DOIs | |
Publication status | Published - 26 Aug 2020 |
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Dive into the research topics of 'The composition and turnover of the Arabidopsis thaliana 80S cytosolic ribosome'. Together they form a unique fingerprint.Projects
- 2 Finished
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Defining factors in the control of protein turnover in plants
Millar, H. (Investigator 01)
ARC Australian Research Council
1/01/18 → 31/03/21
Project: Research
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ARC Centre of Excellence in Plant Energy Biology 2014 (CPEB2)
Millar, H. (Investigator 01), Pogson, B. (Investigator 02), Tyerman, S. (Investigator 03), Small, I. (Investigator 04), Whelan, J. (Investigator 05), Borevitz, J. (Investigator 06), Lister, R. (Investigator 07), Atkin, O. (Investigator 08) & Munns, R. (Investigator 09)
ARC Australian Research Council
1/01/14 → 31/05/21
Project: Research