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The role of oxidative stress in inhibiting FOXO1 through activating Cdk5 in neuronal damage / 西安交通大学学报(医学版)
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wpr-844021
Biblioteca responsável: WPRO
ABSTRACT

Objective:

To investigate the role of oxidative stress in inhibiting forkhead box O1 (FOXO1) through activating cyclin-dependent kinase 5 (Cdk5) in neuronal damage.

Methods:

Cdk5 was coexpressed with p39 in HEK cells together with wild-type FOXO1 or a nonphosphorylatable FOXO1-S249A mutant; FOXO1 phosphorylation was immunoblotted through pS249-FOXO1 antibody which specific recognized FOXO1 Ser249 phosphorylation. Cdk5 or p39 was coexpressed in HEK cells together with FOXO1 and their interaction was explored through Co-IP. In primary cortical culture neurons, various Cdk5 activators (p25, p35, and p39) were overexpressed together with Cdk5. FOXO1 transcriptional activity was tested by luciferase assay. We detected the effect of Cdk5 inhibitor roscovitine on FOXO1 transcriptional activity in primary neurons. We compared FOXO1 subcellular localization of wild type with Cdk5 heterozygous mice cortical brain tissues through cytoplasmic and nuclear extracts. In addition, we administered oxidative stress (H2O2 and glutamate treatment) in primary neurons and examined the effect on the cdk5 activity by Western blot, FOXO1 transcriptional activity by luciferase assay, and neuronal damage through stained caspase3.

Results:

① Identification of Cdk5/p39 was capable of phosphorylating the transcription factor FOXO1 at Ser249, and associated with it. ② Activated Cdk5 dramatically inhibited FOXO1 transcriptional activity. In contrast, roscovitine, Cdk5 inhibitor, significantly increased FOXO1 transcriptional activity and led to FOXO1 translocation into the nucleus. ③ FOXO1 protein was found predominately in the nucleus in the Cdk5 heterozygous mice. H2O2 or glutamate treatment promoted p35 to be cleaved to p25, and inhibited FOXO1 transcriptional activity. Importantly, this effect of oxidative stress was abolished when Cdk5 was silenced by shRNA. ⑤ Oxidative stress dramatically increased caspase3 immunostaining in primary neurons.

Conclusion:

Cdk5/p39 inhibits FOXO1 activity through phosphorylation and association. Moreover, oxidative stress inhibits FOXO1 activity through activating Cdk5, and leads to neuronal death.

Texto completo: Disponível Base de dados: WPRIM (Pacífico Ocidental) Tipo de estudo: Estudo prognóstico Idioma: Chinês Revista: Journal of Xi'an Jiaotong University(Medical Sciences) Ano de publicação: 2019 Tipo de documento: Artigo
Texto completo: Disponível Base de dados: WPRIM (Pacífico Ocidental) Tipo de estudo: Estudo prognóstico Idioma: Chinês Revista: Journal of Xi'an Jiaotong University(Medical Sciences) Ano de publicação: 2019 Tipo de documento: Artigo
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