Post-translational regulation of Ascl1 during neurogenesis

Detalhes bibliográficos
Autor(a) principal: Marques, Raquel Forte
Data de Publicação: 2021
Tipo de documento: Dissertação
Idioma: eng
Título da fonte: Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
Texto Completo: http://hdl.handle.net/10451/53896
Resumo: Tese de mestrado, Biologia Evolutiva e do Desenvolvimento, Universidade de Lisboa, Faculdade de Ciências, 2022
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spelling Post-translational regulation of Ascl1 during neurogenesisFosforilaçãoO-glicosilaçãoDiferenciação NeuronalCarga das proteinasLigação á cromatinaTeses de mestrado - 2022Departamento de Biologia AnimalTese de mestrado, Biologia Evolutiva e do Desenvolvimento, Universidade de Lisboa, Faculdade de Ciências, 2022Neurogenesis in vertebrates is to a large extent regulated by proneural transcriptional factors (TFs) such as Ascl1. Ascl1 is expressed in the germinal layers of the ventral telencephalon, promoting sequentially the proliferation and differentiation of neural stem/progenitors throughout the neuronal lineage. Ac cording to the current model an increase in Ascl1 activity determines the onset of neuronal differentia tion. Multisite phosphorylation of proneural TFs in Serine/Proline (SP) sites outside the DNA-binding domain has been proposed to decrease neurogenic activity, by changing their overall electrostatic po tential. O-GlcNAcylation, a neutrally charged PTM catalyzed by O-GlcNAc transferase OGT, can counteract phosphorylation by targeting the same residues. Few studies have implicated O-GlcNAcyl ation in neurogenesis regulation, but the exact mechanisms (and how this may impact Ascl1 activity) is not known. Here, using western-blot analysis we show for the first time that endogenous Ascl1 protein is highly phosphorylated in neural stem/progenitors in culture, and in the embryonic ventral telencephalon. Additionally, we observe a decrease in Ascl1 phosphorylation during neuronal differentiation. Preliminary results, using overexpression of a phosphodead version of Ascl1 (with six SP sites mutated), suggest multisite phosphorylation does not impact protein stability, but results in an increase in its cytoplasmic and chromatin bound fractions. This is in line with the possibility that multisite phosphorylation decreases Ascl1 activity, by reducing its association with negatively charged chromatin. Furthermore, we used immunofluorescence and western-blot analysis to characterize O GlcNAcylation in cultured neural stem/progenitor cells. We found O-GlcNAc is predominantly nuclear, becoming even more nuclear during neuronal differentiation. Unexpectedly, we found that a pharmacological inhibitor of O-GlcNAc transferase (OGT) drastically reduces Ascl1 protein levels in cultured neural stem/progenitor cells. Additionally, co-expression of OGT with Ascl1 (tethered to its heterodimeric partner E47) indicates Ascl1/E47 heterodimers can be targeted by O-GlcNAcylation. This suggests different mechanisms by which O-GlcNAcylation may regulate Ascl1, which should be further investigated.Castro, Diogo S.Thorsteinsdóttir, Sólveig, 1962-Repositório da Universidade de LisboaMarques, Raquel Forte202220212024-12-30T00:00:00Z2022-01-01T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisapplication/pdfhttp://hdl.handle.net/10451/53896enginfo:eu-repo/semantics/embargoedAccessreponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãoinstacron:RCAAP2023-11-08T17:00:08Zoai:repositorio.ul.pt:10451/53896Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-19T22:04:52.992447Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãofalse
dc.title.none.fl_str_mv Post-translational regulation of Ascl1 during neurogenesis
title Post-translational regulation of Ascl1 during neurogenesis
spellingShingle Post-translational regulation of Ascl1 during neurogenesis
Marques, Raquel Forte
Fosforilação
O-glicosilação
Diferenciação Neuronal
Carga das proteinas
Ligação á cromatina
Teses de mestrado - 2022
Departamento de Biologia Animal
title_short Post-translational regulation of Ascl1 during neurogenesis
title_full Post-translational regulation of Ascl1 during neurogenesis
title_fullStr Post-translational regulation of Ascl1 during neurogenesis
title_full_unstemmed Post-translational regulation of Ascl1 during neurogenesis
title_sort Post-translational regulation of Ascl1 during neurogenesis
author Marques, Raquel Forte
author_facet Marques, Raquel Forte
author_role author
dc.contributor.none.fl_str_mv Castro, Diogo S.
Thorsteinsdóttir, Sólveig, 1962-
Repositório da Universidade de Lisboa
dc.contributor.author.fl_str_mv Marques, Raquel Forte
dc.subject.por.fl_str_mv Fosforilação
O-glicosilação
Diferenciação Neuronal
Carga das proteinas
Ligação á cromatina
Teses de mestrado - 2022
Departamento de Biologia Animal
topic Fosforilação
O-glicosilação
Diferenciação Neuronal
Carga das proteinas
Ligação á cromatina
Teses de mestrado - 2022
Departamento de Biologia Animal
description Tese de mestrado, Biologia Evolutiva e do Desenvolvimento, Universidade de Lisboa, Faculdade de Ciências, 2022
publishDate 2021
dc.date.none.fl_str_mv 2021
2022
2022-01-01T00:00:00Z
2024-12-30T00:00:00Z
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/masterThesis
format masterThesis
status_str publishedVersion
dc.identifier.uri.fl_str_mv http://hdl.handle.net/10451/53896
url http://hdl.handle.net/10451/53896
dc.language.iso.fl_str_mv eng
language eng
dc.rights.driver.fl_str_mv info:eu-repo/semantics/embargoedAccess
eu_rights_str_mv embargoedAccess
dc.format.none.fl_str_mv application/pdf
dc.source.none.fl_str_mv reponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação
instacron:RCAAP
instname_str Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação
instacron_str RCAAP
institution RCAAP
reponame_str Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
collection Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
repository.name.fl_str_mv Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação
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