Journal
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
Volume 23, Issue 7, Pages -Publisher
MDPI
DOI: 10.3390/ijms23073646
Keywords
SYNGR3; synaptogyrin; synaptic dysfunction; Parkinson's disease; gene regulation; promoter analysis; cis-regulatory elements; NURR1; transactivator
Funding
- Tai Hung Fai Charitable Foundation-Edwin S H Leong Research Programme for Parkinson's Disease
- Henry G. Leong Endowed Professorship in Neurology
- University Grant Committee (UGC), University of Hong Kong, Hong Kong SAR, China
- Hong Kong PhD Fellowship Scheme (HKPF), Research Grant Council, Hong Kong SAR, China
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This study investigated whether NURR1 can regulate the expression of SYNGR3. In silico analysis and experimental validation revealed that NURR1 can bind to the cis-acting sites of the SYNGR3 gene and activation of NURR1 can increase the expression of SYNGR3. This finding suggests that inducing SYNGR3 expression by activating NURR1 may be a potential therapeutic strategy to attenuate synaptic dysfunction in Parkinson's disease.
Synaptogyrin-3 (SYNGR3) is a synaptic vesicular membrane protein. Amongst four homologues (SYNGR1 to 4), SYNGR1 and 3 are especially abundant in the brain. SYNGR3 interacts with the dopamine transporter (DAT) to facilitate dopamine (DA) uptake and synaptic DA turnover in dopaminergic transmission. Perturbed SYNGR3 expression is observed in Parkinson's disease (PD). The regulatory elements which affect SYNGR3 expression are unknown. Nuclear-receptor-related-1 protein (NURR1) can regulate dopaminergic neuronal differentiation and maintenance via binding to NGFI-B response elements (NBRE). We explored whether NURR1 can regulate SYNGR3 expression using an in silico analysis of the 5 '-flanking region of the human SYNGR3 gene, reporter gene activity and an electrophoretic mobility shift assay (EMSA) of potential cis-acting sites. In silico analysis of two genomic DNA segments (1870 bp 5 '-flanking region and 1870 + 159 bp of first exon) revealed one X Core Promoter Element 1 (XCPE1), two SP1, and three potential non-canonical NBRE response elements (ncNBRE) but no CAAT or TATA box. The longer segment exhibited gene promoter activity in luciferase reporter assays. Site-directed mutagenesis of XCPE1 decreased promoter activity in human neuroblastoma SH-SY5Y (down arrow 43.2%) and human embryonic kidney HEK293 cells (down arrow 39.7%). EMSA demonstrated NURR1 binding to these three ncNBRE. Site-directed mutagenesis of these ncNBRE reduced promoter activity by 11-17% in SH-SY5Y (neuronal) but not in HEK293 (non-neuronal) cells. C-DIM12 (Nurr1 activator) increased SYNGR3 protein expression in SH-SY5Y cells and its promoter activity using a real-time luciferase assay. As perturbed vesicular function is a feature of major neurodegenerative diseases, inducing SYNGR3 expression by NURR1 activators may be a potential therapeutic target to attenuate synaptic dysfunction in PD.
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