2026-04-17T20:17:35Zhttps://recercat.cat/oai/request

oai:recercat.cat:10230/579502025-12-12T04:08:24Zcom_2072_6col_2072_452952

Dynamic interplay between RPL3- and RPL3L-containing ribosomes modulates mitochondrial activity in the mammalian heart Milenkovic, Ivan Santos Vieira, Helaine Graziele Lucas, Morghan C. Ruiz-Orera, Jorge Patone, Giannino Kesteven, Scott Wu, Jianxin Feneley, Michael Espadas, Guadalupe Sabidó Aguadé, Eduard, 1981- Hübner, Norbert van Heesch, Sebastiaan Völkers, Mirko Novoa, Eva Maria The existence of naturally occurring ribosome heterogeneity is now a well-acknowledged phenomenon. However, whether this heterogeneity leads to functionally diverse 'specialized ribosomes' is still a controversial topic. Here, we explore the biological function of RPL3L (uL3L), a ribosomal protein (RP) paralogue of RPL3 (uL3) that is exclusively expressed in skeletal muscle and heart tissues, by generating a viable homozygous Rpl3l knockout mouse strain. We identify a rescue mechanism in which, upon RPL3L depletion, RPL3 becomes up-regulated, yielding RPL3-containing ribosomes instead of RPL3L-containing ribosomes that are typically found in cardiomyocytes. Using both ribosome profiling (Ribo-seq) and a novel orthogonal approach consisting of ribosome pulldown coupled to nanopore sequencing (Nano-TRAP), we find that RPL3L modulates neither translational efficiency nor ribosome affinity towards a specific subset of transcripts. In contrast, we show that depletion of RPL3L leads to increased ribosome-mitochondria interactions in cardiomyocytes, which is accompanied by a significant increase in ATP levels, potentially as a result of fine-tuning of mitochondrial activity. Our results demonstrate that the existence of tissue-specific RP paralogues does not necessarily lead to enhanced translation of specific transcripts or modulation of translational output. Instead, we reveal a complex cellular scenario in which RPL3L modulates the expression of RPL3, which in turn affects ribosomal subcellular localization and, ultimately, mitochondrial activity. The European Union's Horizon 2020 Research and Innovation Program under the Marie Skodowska-Curie grant agreement [713673]; the Australian Research Council [DE170100506 to E.M.N.]; the Spanish Ministry of Economy, Industry and Competitiveness (MEIC) [PGC2018-098152-A-100 to E.M.N.]; the European Union Horizon 2020 Research and Innovation Program ERC advanced grant [AdG788970 to N.H.] and ERC starting grant [StG101042103 to E.M.N.]; the Leducq Foundation [16CVD03 to N.H.]; the Chan Zuckerberg Foundation [2019-20266 to N.H.]; and ‘la Caixa’ INPhINIT PhD fellowship [LCF/BQ/DI18/11660028 to I.M.]. We acknowledge the support of the MEIC to the EMBL partnership, Centro de Excelencia Severo Ochoa and CERCA Programme/Generalitat de Catalunya. The CRG/UPF Proteomics Unit is part of the Spanish Infrastructure for Omics Technologies (ICTS OmicsTech) and it is a member of the ProteoRed PRB3 consortium which is supported by grant PT17/0019 of the PE I + D + i 2013-2016 from the Instituto de Salud Carlos III (ISCIII) and ERDF. Conflict of interest statement. E.M.N. has received travel expenses from ONT to participate in nanopore conferences. I.M. has received a travel bursary from ONT to present his work in international conferences. E.M.N. is Scientific Advisory Board member for IMMAGINA Biotech. The authors declare that they have no competing interests. 2023-09-26T06:30:54Z 2023-09-26T06:30:54Z 2023 info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion Nucleic Acids Res. 2023;51(11):5301-24 info:eu-repo/grantAgreement/EC/H2020/713673 info:eu-repo/grantAgreement/ES/2PE/PGC2018-098152-A-100 info:eu-repo/grantAgreement/EC/H2020/788970 © The Author(s) 2023. This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial License (http://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com http://creativecommons.org/licenses/by-nc/4.0/ info:eu-repo/semantics/openAccess Oxford University Press