Bioinformatic and functional analysis of promoter region of human SLC25A13 gene
| Autor: | Bing-Xiao Li, Jun-Lin Chen, Qing-Hua Zhou, Zhen Cai, Zhan-Hui Zhang |
|---|---|
| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
TATA box CAAT box Organic Anion Transporters Mitochondrial Membrane Transport Proteins 03 medical and health sciences 0302 clinical medicine Initiator element Transcription (biology) Genetics Humans Promoter Regions Genetic Gene Reporter gene biology Base Sequence Calcium-Binding Proteins Computational Biology Promoter General Medicine Molecular biology 030104 developmental biology Citrin 030220 oncology & carcinogenesis Mutation biology.protein |
| Zdroj: | Gene. 693 |
| ISSN: | 1879-0038 |
| Popis: | The human SLC25A13 gene encodes the liver type aspartate/glutamate carrier isoform 2 (AGC2, commonly named as citrin), which plays a key role in the main NADH-shuttle of human hepatocyte. Biallelic SLC25A13 mutations result in Citrin deficiency (CD). In order to identify the important regulatory region of SLC25A13 gene and elucidate the way how potential promoter mutations affect the citrin expression, we performed promoter deletion analysis and established the reporter constructs of luciferase gene-carrying SLC25A13 promoter containing several mutations located in putative transcription factor-binding sites. The luciferase activities of all promoter constructs were measured using a Dual-Luciferase Reporter Assay System. Bioinformatic analysis showed that the promoter of SLC25A13 gene lacks TATA box and obviously typical initiator element, but contains a CCAAT box and two GC box. Promoter deletion analysis confirmed the region from -221 to -1 upstream ATG was essential for SLC25A13 to maintain the promoter activity. We utilized dual-luciferase reporter system as function analytical model to tentatively assess the effect of artificially constructed promoter mutations on citrin expression, and our analysis revealed that mutated putative CCAAT box and GC box could significantly affect the citrin expression. Our study confirmed the important SLC25A13 promoter regions that influenced citrin expression in HL7702 cells, and constructed a function analytical model. This work may be useful to further identify the pathogenic mutations leading to CD in the promoter region. |
| Databáze: | OpenAIRE |
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