Molecular cloning and characterization of the oxidative stress sensitive kinase PKD1 gene promoter in a cell culture model of Parkinson's disease
Başlık çevirisi mevcut değil.
- Tez No: 403135
- Danışmanlar: Dr. ANUMANTHA G. KANTHASAMY
- Tez Türü: Yüksek Lisans
- Konular: Genetik, Genetics
- Anahtar Kelimeler: Belirtilmemiş.
- Yıl: 2011
- Dil: İngilizce
- Üniversite: Iowa State University
- Enstitü: Yurtdışı Enstitü
- Ana Bilim Dalı: Belirtilmemiş.
- Bilim Dalı: Belirtilmemiş.
- Sayfa Sayısı: 87
Özet
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Özet (Çeviri)
Parkinson's disease (PD) is the second most common neurodegenerative disorder characterized by selective loss of dopaminergic neurons in the substantia nigra. Oxidative stress-induced apoptosis has been strongly implicated in the pathogenesis of PD. Recently, we demonstrated that protein kinase D1 (PKD1) protects dopaminergic neurons against oxidative stress-induced apoptosis. Even though the activation of PKD1 has been widely investigated, the transcriptional regulation of PKD1 remains largely unknown. In order to investigate the transcriptional regulation of PKD1, we cloned and characterized the 5'- flanking region (1620 bp) of the mouse PKD1 gene. Progressive 5' and 3' deletion analyses revealed that the -250/+113 promoter region contains the full promoter activity in transiently transfected MN9D dopaminergic neuronal cells and that the noncoding part of exon1 harbors both positive and negative regulatory elements. Overexpression of Sp1, Sp3, and NF-kB p65 proteins stimulated PKD1 promoter activity. Moreover, overexpression of CREB binding protein (CBP) enhanced Sp3-mediated transactivation of the PKD1 promoter. Both PKD1 promoter activity and endogenous PKD1 mRNA in MN9D cells were attenuated by the sp inhibitor mithramycin A. Furthermore, we demonstrated that the histone deacetylase (HDAC) inhibitors trichostatin A (TSA) and sodium butyrate (NaBu) and the DNA demethylating agent 5-Aza-2'-deoxycytidine (5-aza-dC) can upregulate PKD1 mRNA expression in MN9D dopaminergic neuronal cells. Collectively, our findings suggest that Sp3, the major transcriptional activator of the PKD1 gene, and epigenetic mechanisms such as DNA methylation and histone modifications, play essential roles in regulating the expression of the PKD1 gene in dopaminergic neuronal cells.
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