Publicação
Investigating the role of ELP3 overexpression in cellular models of Alzheimer’s disease
| Resumo: | Alzheimer's disease (AD) is a neurodegenerative disorder characterized by severe cognitive decline. However, the molecular mechanisms underlying AD onset and progression remain unclear. Recently, tRNA modifications catalyzed by different tRNA modifying enzymes have emerged as key regulators of translation, essential for accurate mRNA decoding. Disruption of tRNA modifications have been found in several neurological disorders and we have recently shown that the expression of ELP3, a tRNA-modifying enzyme, and the levels of its dependent wobble uridine tRNA modifications are reduced in AD. In this study, we investigated the effect of ELP3 overexpression in AD cellular models, namely SH-SY5Y-appSwe (SH-SWE) cells when compared to SH-SY5Y expressing wild-type APP (SH-WT). We employed a plasmid transfection protocol containing an ELP3-expression plasmid (pELP3) and an empty plasmid (pMOCK), serving as a control. ELP3 expression, 2-thiolation levels, insoluble fraction abundance, APP levels, global protein synthesis, and proteostasis markers were analyzed after plasmid transfection. Although 2-thiolation levels increased in SH-WT upon ELP3 overexpression as expected, the opposite was observed in in SH-SWE cells. Moreover, overexpression of ELP3 in SH-SWE cells led to higher insoluble protein levels, reduced global translation, and no recovery of proteostasis markers. Overall, our results indicate that ELP3 overexpression is not a good strategy to recover the basal stress levels to which AD cells are subjected to, but may represent a valid strategy to alleviate stress in non-diseased cells, constituting a putative strategy to alleviate aging-related translation impairments. |
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| Autores principais: | Lacerda, Julia Costa de |
| Assunto: | Alzheimer’s disease tRNA modifications tRNA-modifying enzymes ELP3 Translation |
| Ano: | 2025 |
| País: | Portugal |
| Tipo de documento: | dissertação de mestrado |
| Tipo de acesso: | acesso embargado |
| Instituição associada: | Universidade de Aveiro |
| Idioma: | inglês |
| Origem: | RIA - Repositório Institucional da Universidade de Aveiro |
| Resumo: | Alzheimer's disease (AD) is a neurodegenerative disorder characterized by severe cognitive decline. However, the molecular mechanisms underlying AD onset and progression remain unclear. Recently, tRNA modifications catalyzed by different tRNA modifying enzymes have emerged as key regulators of translation, essential for accurate mRNA decoding. Disruption of tRNA modifications have been found in several neurological disorders and we have recently shown that the expression of ELP3, a tRNA-modifying enzyme, and the levels of its dependent wobble uridine tRNA modifications are reduced in AD. In this study, we investigated the effect of ELP3 overexpression in AD cellular models, namely SH-SY5Y-appSwe (SH-SWE) cells when compared to SH-SY5Y expressing wild-type APP (SH-WT). We employed a plasmid transfection protocol containing an ELP3-expression plasmid (pELP3) and an empty plasmid (pMOCK), serving as a control. ELP3 expression, 2-thiolation levels, insoluble fraction abundance, APP levels, global protein synthesis, and proteostasis markers were analyzed after plasmid transfection. Although 2-thiolation levels increased in SH-WT upon ELP3 overexpression as expected, the opposite was observed in in SH-SWE cells. Moreover, overexpression of ELP3 in SH-SWE cells led to higher insoluble protein levels, reduced global translation, and no recovery of proteostasis markers. Overall, our results indicate that ELP3 overexpression is not a good strategy to recover the basal stress levels to which AD cells are subjected to, but may represent a valid strategy to alleviate stress in non-diseased cells, constituting a putative strategy to alleviate aging-related translation impairments. |
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