Functional characterization of the neuron-restrictive silencer element in the human tryptophan hydroxylase 2 gene expression
Autor: | Masayuki Oda, Luca Colucci-D'Amato, Masaaki Tsubonoya, Tomoko Hiroi, Yukino Nawa, Hanae Kaneko, Maria Teresa Gentile, Hiroaki Matsui, Ryoya Takahashi |
---|---|
Přispěvatelé: | Nawa, Yukino, Kaneko, Hanae, Oda, Masayuki, Tsubonoya, Masaaki, Hiroi, Tomoko, Gentile, Maria Teresa, COLUCCI D'AMATO, Generoso Luca, Takahashi, Ryoya, Matsui, Hiroaki |
Rok vydání: | 2017 |
Předmět: |
0301 basic medicine
Gene knockdown promoter TPH1 TPH2 Tryptophan hydroxylase Biology Biochemistry Molecular biology 03 medical and health sciences Cellular and Molecular Neuroscience 030104 developmental biology 0302 clinical medicine NRSF Gene expression gene expression Transcriptional regulation serotonergic neuron transcriptional regulation Histone deacetylase Psychological repression 030217 neurology & neurosurgery |
Zdroj: | Journal of Neurochemistry. 142:827-840 |
ISSN: | 0022-3042 |
Popis: | Tryptophan hydroxylase 2 (TPH2) is the key enzyme in the synthesis of neuronal serotonin. Although previous studies suggest that TPH2 neuron-restrictive silencer element (NRSE) functions as a negative regulator dependent on neuron-restrictive silencer factor (NRSF) activity, the underlying mechanisms are yet to be fully elucidated. Here, we show a detailed analysis of the NRSE-mediated repression of the human TPH2 (hTPH2) promoter activity in RN46A cells, a cell line derived from rat raphe neurons. Quantitative real-time RT-PCR analysis revealed the expression of serotonergic marker genes (Mash1, Nkx2.2, Gata2, Gata3, Lmx1b, Pet-1, 5-Htt, and Vmat2) and Nrsf gene in RN46A cells. Tph1 mRNA is the prevalent form expressed in RN46A cells; Tph2 mRNA is also expressed but at a lower level. Electrophoretic mobility shift assays and reporter assays showed that hTPH2 NRSE is necessary for the efficient DNA binding of NRSF and for the NRSF-dependent repression of the hTPH2 promoter activity. The hTPH2 promoter activity was increased by knockdown of NRSF, or over-expression of the engineered NRSF (a dominant-negative mutant or a DNA-binding domain and activation domain fusion protein). MS-275, a class I histone deacetylase (HDAC) inhibitor, was found to be more potent than MC-1568, a class II HDAC inhibitor, in enhancing the hTPH2 promoter activity. Furthermore, treatment with the ubiquitin-specific protease 7 deubiquitinase inhibitors, P-22077 or HBX 41108, increased the hTPH2 promoter activity. Collectively, our data demonstrate that the hTPH2 NRSE-mediated promoter repression via NRSF involves class I HDACs and is modulated by the ubiquitin-specific protease 7-mediated deubiquitination and stabilization of NRSF. |
Databáze: | OpenAIRE |
Externí odkaz: |