Ethanol-Induced Upregulation of 10-Formyltetrahydrofolate Dehydrogenase Helps Relieve Ethanol-Induced Oxidative Stress
| Autor: | Bing Hung Chen, Wen Ni Chang, Yi Shao Chung, Tseng Ting Kao, Gang Hui Lee, Chia Jen Lin, Tzu Fun Fu, Jan Jong Hung, Tsun Hsien Hsiao |
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| Jazyk: | angličtina |
| Rok vydání: | 2014 |
| Předmět: |
Embryo
Nonmammalian Molecular Sequence Data Dehydrogenase Biology medicine.disease_cause Folic Acid Downregulation and upregulation medicine CCAAT-Enhancer-Binding Protein-alpha Animals Promoter Regions Genetic Molecular Biology Zebrafish Tetrahydrofolates chemistry.chemical_classification Messenger RNA Oxidoreductases Acting on CH-NH Group Donors Binding Sites Base Sequence Ethanol Mutagenesis Central Nervous System Depressants Gene Expression Regulation Developmental Cell Biology Articles Zebrafish Proteins biology.organism_classification Molecular biology Up-Regulation Oxidative Stress Enzyme chemistry Gene Knockdown Techniques Mutagenesis Site-Directed Oxidative stress Intracellular |
| Popis: | Alcoholism induces folate deficiency and increases the risk for embryonic anomalies. However, the interplay between ethanol exposure and embryonic folate status remains unclear. To investigate how ethanol exposure affects embryonic folate status and one-carbon homeostasis, we incubated zebrafish embryos in ethanol and analyzed embryonic folate content and folate enzyme expression. Exposure to 2% ethanol did not change embryonic total folate content but increased the tetrahydrofolate level approximately 1.5-fold. The expression of 10-formyltetrahydrofolate dehydrogenase (FDH), a potential intracellular tetrahydrofolate reservoir, was increased in both mRNA and protein levels. Overexpressing recombinant FDH in embryos alleviated the ethanol-induced oxidative stress in ethanol-exposed embryos. Further characterization of the zebrafish fdh promoter revealed that the -124/+40 promoter fragment was the minimal region required for transactivational activity. The results of site-directed mutagenesis and binding analysis revealed that Sp1 is involved in the basal level of expression of fdh but not in ethanol-induced upregulation of fdh. On the other hand, CEBPα was the protein that mediated the ethanol-induced upregulation of fdh, with an approximately 40-fold increase of fdh promoter activity when overexpressed in vitro. We concluded that upregulation of fdh involving CEBPα helps relieve embryonic oxidative stress induced by ethanol exposure. |
| Databáze: | OpenAIRE |
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