Arginyltransferase 1 modulates p62-driven autophagy via mTORC1/AMPk signaling
Fil: Bonnet Laura V. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Departamento de Química Biológica Ranwel Caputto, Argentina.
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| Format: | info:eu-repo/semantics/publishedVersion |
| Language: | eng |
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BioMed Central Ltd. Part of Springer Nature
2024
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| Online Access: | http://hdl.handle.net/11086/552355 https://link.springer.com/article/10.1186/s12964-024-01499-9 https://doi.org/10.1186/s12964-024-01499-9 |
| _version_ | 1802926365383589888 |
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| author | Bonnet, Laura V. Palandri, Anabela Flores Martin, Jesica B. Hallak, Marta E. |
| author2 | https://orcid.org/0000-0001-5493-0949 |
| author_facet | https://orcid.org/0000-0001-5493-0949 Bonnet, Laura V. Palandri, Anabela Flores Martin, Jesica B. Hallak, Marta E. |
| author_sort | Bonnet, Laura V. |
| collection | Repositorio Digital Universitario |
| description | Fil: Bonnet Laura V. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Departamento de Química Biológica Ranwel Caputto, Argentina. |
| format | info:eu-repo/semantics/publishedVersion |
| id | rdu-unc.552355 |
| institution | Universidad Nacional de Cordoba |
| language | eng |
| publishDate | 2024 |
| publisher | BioMed Central Ltd. Part of Springer Nature |
| record_format | dspace |
| spelling | rdu-unc.5523552024-06-23T06:22:46Z Arginyltransferase 1 modulates p62-driven autophagy via mTORC1/AMPk signaling Bonnet, Laura V. Palandri, Anabela Flores Martin, Jesica B. Hallak, Marta E. https://orcid.org/0000-0001-5493-0949 https://orcid.org/0000-0003-4856-879X https://orcid.org/0000-0002-1619-399X Arginylation Arginyltransferase 1 Autophagy mTORC1 Posttranslational modifcation p62/SQSTM1 info:eu-repo/semantics/publishedVersion Fil: Bonnet Laura V. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Departamento de Química Biológica Ranwel Caputto, Argentina. Fil: Bonnet Laura V. Consejo Nacional de Investigaciones Científicas y Técnicas. Argentina. Centro de Investigaciones en Química Biológica de Córdoba, Argentina. Fil: Palandri Anabela. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Departamento de Química Biológica Ranwel Caputto, Argentina. Fil: Palandri Anabela. Consejo Nacional de Investigaciones Científicas y Técnicas. Argentina. Centro de Investigaciones en Química Biológica de Córdoba, Argentina. Fil: Flores Martin Jesica B. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Departamento de Química Biológica Ranwel Caputto, Argentina. Fil: Flores Martin Jesica B. Consejo Nacional de Investigaciones Científicas y Técnicas. Argentina. Centro de Investigaciones en Química Biológica de Córdoba, Argentina. Fil: Hallak Marta E. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Departamento de Química Biológica Ranwel Caputto, Argentina. Fil: Hallak Marta E. Consejo Nacional de Investigaciones Científicas y Técnicas. Argentina. Centro de Investigaciones en Química Biológica de Córdoba, Argentina. Background. Arginyltransferase (Ate1) orchestrates posttranslational protein arginylation, a pivotal regulator of cellular proteolytic processes. In eukaryotic cells, two interconnected systems—the ubiquitin proteasome system (UPS) and macroautophagy—mediate proteolysis and cooperate to maintain quality protein control and cellular homeostasis. Previous studies have shown that N-terminal arginylation facilitates protein degradation through the UPS. Dysregulation of this machinery triggers p62-mediated autophagy to ensure proper substrate processing. Nevertheless, how Ate1 operates through this intricate mechanism remains elusive. Methods. We investigated Ate1 subcellular distribution through confocal microscopy and biochemical assays using cells transiently or stably expressing either endogenous Ate1 or a GFP-tagged Ate1 isoform transfected in CHO-K1 or MEFs, respectively. To assess Ate1 and p62-cargo clustering, we analyzed their colocalization and multimerization status by immunofluorescence and nonreducing immunoblotting, respectively. Additionally, we employed Ate1 KO cells to examine the role of Ate1 in autophagy. Ate1 KO MEFs cells stably expressing GFP-tagged Ate1-1 isoform were used as a model for phenotype rescue. Autophagy dynamics were evaluated by analyzing LC3B turnover and p62/SQSTM1 levels under both steady-state and serum-starvation conditions, through immunoblotting and immunofluorescence. We determined mTORC1/AMPk activation by assessing mTOR and AMPk phosphorylation through immunoblotting, while mTORC1 lysosomal localization was monitored by confocal microscopy. Results. Here, we report a multifaceted role for Ate1 in the autophagic process, wherein it clusters with p62, facilitates autophagic clearance, and modulates its signaling. Mechanistically, we found that cell-specific inactivation of Ate1 elicits overactivation of the mTORC1/AMPk signaling hub that underlies a failure in autophagic flux and subsequent substrate accumulation, which is partially rescued by ectopic expression of Ate1. Statistical significance was assessed using a two-sided unpaired t test with a significance threshold set at P<0.05. Conclusions. Our findings uncover a critical housekeeping role of Ate1 in mTORC1/AMPk-regulated autophagy, as a potential therapeutic target related to this pathway, that is dysregulated in many neurodegenerative and cancer diseases. info:eu-repo/semantics/publishedVersion Fil: Bonnet Laura V. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Departamento de Química Biológica Ranwel Caputto, Argentina. Fil: Bonnet Laura V. Consejo Nacional de Investigaciones Científicas y Técnicas. Argentina. Centro de Investigaciones en Química Biológica de Córdoba, Argentina. Fil: Palandri Anabela. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Departamento de Química Biológica Ranwel Caputto, Argentina. Fil: Palandri Anabela. Consejo Nacional de Investigaciones Científicas y Técnicas. Argentina. Centro de Investigaciones en Química Biológica de Córdoba, Argentina. Fil: Flores Martin Jesica B. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Departamento de Química Biológica Ranwel Caputto, Argentina. Fil: Flores Martin Jesica B. Consejo Nacional de Investigaciones Científicas y Técnicas. Argentina. Centro de Investigaciones en Química Biológica de Córdoba, Argentina. Fil: Hallak Marta E. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Departamento de Química Biológica Ranwel Caputto, Argentina. Fil: Hallak Marta E. Consejo Nacional de Investigaciones Científicas y Técnicas. Argentina. Centro de Investigaciones en Química Biológica de Córdoba, Argentina. 2024-06-23T03:14:48Z 2024-06-23T03:14:48Z 2024-01-31 article Bonnet, L.V., Palandri, A., Flores-Martin, J.B. et al. Arginyltransferase 1 modulates p62-driven autophagy via mTORC1/AMPk signaling. Cell Commun Signal 22, 87 (2024). https://doi.org/10.1186/s12964-024-01499-9 http://hdl.handle.net/11086/552355 1478-811X https://link.springer.com/article/10.1186/s12964-024-01499-9 https://doi.org/10.1186/s12964-024-01499-9 eng Attribution-NonCommercial-NoDerivatives 4.0 International http://creativecommons.org/licenses/by-nc-nd/4.0/ BioMed Central Ltd. Part of Springer Nature |
| spellingShingle | Arginylation Arginyltransferase 1 Autophagy mTORC1 Posttranslational modifcation p62/SQSTM1 Bonnet, Laura V. Palandri, Anabela Flores Martin, Jesica B. Hallak, Marta E. Arginyltransferase 1 modulates p62-driven autophagy via mTORC1/AMPk signaling |
| title | Arginyltransferase 1 modulates p62-driven autophagy via mTORC1/AMPk signaling |
| title_full | Arginyltransferase 1 modulates p62-driven autophagy via mTORC1/AMPk signaling |
| title_fullStr | Arginyltransferase 1 modulates p62-driven autophagy via mTORC1/AMPk signaling |
| title_full_unstemmed | Arginyltransferase 1 modulates p62-driven autophagy via mTORC1/AMPk signaling |
| title_short | Arginyltransferase 1 modulates p62-driven autophagy via mTORC1/AMPk signaling |
| title_sort | arginyltransferase 1 modulates p62 driven autophagy via mtorc1 ampk signaling |
| topic | Arginylation Arginyltransferase 1 Autophagy mTORC1 Posttranslational modifcation p62/SQSTM1 |
| url | http://hdl.handle.net/11086/552355 https://link.springer.com/article/10.1186/s12964-024-01499-9 https://doi.org/10.1186/s12964-024-01499-9 |
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