Depletion of <i>SNORA33</i> Abolishes ψ of 28S-U4966 and Affects the Ribosome Translational Apparatus


Chabronova, Alzbeta ORCID: 0000-0002-9733-444X, van den Akker, Guus, Housmans, Bas AC, Caron, Marjolein MJ, Cremers, Andy, Surtel, Don AM, Peffers, Mandy J ORCID: 0000-0001-6979-0440, van Rhijn, Lodewijk W, Marchand, Virginie, Motorin, Yuri et al (show 1 more authors) (2023) Depletion of <i>SNORA33</i> Abolishes ψ of 28S-U4966 and Affects the Ribosome Translational Apparatus. INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES, 24 (16). 12578-.

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Abstract

Eukaryotic ribosomes are complex molecular nanomachines translating genetic information from mRNAs into proteins. There is natural heterogeneity in ribosome composition. The pseudouridylation (ψ) of ribosomal RNAs (rRNAs) is one of the key sources of ribosome heterogeneity. Nevertheless, the functional consequences of ψ-based ribosome heterogeneity and its relevance for human disease are yet to be understood. Using HydraPsiSeq and a chronic disease model of non-osteoarthritic primary human articular chondrocytes exposed to osteoarthritic synovial fluid, we demonstrated that the disease microenvironment is capable of instigating site-specific changes in rRNA ψ profiles. To investigate one of the identified differential rRNA ψ sites (28S-ψ4966), we generated <i>SNORA22</i> and <i>SNORA33</i> KO SW1353 cell pools using LentiCRISPRv2/Cas9 and evaluated the ribosome translational capacity by <sup>35</sup>S-Met/Cys incorporation, assessed the mode of translation initiation and ribosomal fidelity using dual luciferase reporters, and assessed cellular and ribosomal proteomes by LC-MS/MS. We uncovered that the depletion of <i>SNORA33</i>, but not <i>SNORA22</i>, reduced 28S-ψ4966 levels. The resulting loss of 28S-ψ4966 affected ribosomal protein composition and function and led to specific changes in the cellular proteome. Overall, our pioneering findings demonstrate that cells dynamically respond to disease-relevant changes in their environment by altering their rRNA pseudouridylation profiles, with consequences for ribosome function and the cellular proteome relevant to human disease.

Item Type: Article
Uncontrolled Keywords: ribosomal RNA, 28S, epitranscriptome, ribosome, chondrocytes, osteoarthritis
Divisions: Faculty of Health and Life Sciences
Faculty of Health and Life Sciences > Institute of Life Courses and Medical Sciences
Depositing User: Symplectic Admin
Date Deposited: 31 Aug 2023 09:51
Last Modified: 24 Oct 2023 00:35
DOI: 10.3390/ijms241612578
Related URLs:
URI: https://livrepository.liverpool.ac.uk/id/eprint/3172450
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