Abstract: SA-PO415
Alternative Promoter Type Influences the Expression of the GLA Gene in Human Kidney Cells
Session Information
- Genetic Diseases of the Kidney - III
November 09, 2019 | Location: Exhibit Hall, Walter E. Washington Convention Center
Abstract Time: 10:00 AM - 12:00 PM
Category: Genetic Diseases of the Kidneys
- 1002 Genetic Diseases of the Kidneys: Non-Cystic
Authors
- Al-Obaide, Mohammed A., Texas Tech University Health Sciences Center, Amarillo, Texas, United States
- Vasylyeva, Tetyana L., Texas Tech University Health Sciences Center, Amarillo, Texas, United States
Background
Diagnostic genetic methods for Fabry disease focus on molecular screening for mutations in GLA exons. Intriguingly, alternative promoters are greatly influencing differential gene expression in tissues and malfunctions in many disease-associated genes [Trends Genet (2008) 24(4):167-177; Genome Res (2011) 21(8):1260-1272]. GLA expression is one of more than 50% of human genes that are controlled by alternative promoters. We aimed to investigate the influence of alternative promoters on GLA expression in three types of human kidney cells (embryonic, epithelial and glomerular cells).
Methods
The GLA alternative promoters were searched in TRED, EPD, PrESSto/FANTOM5, and Ensembl databases. RNA extracted from embryonic, epithelial and glomerular kidney cells by RNAzol method. RT q-PCR was used for the analysis of GLA expression. Expression data of GLA alternative promoters were normalized to reference gene HPRT1, and the fold-change in the GLA expression was calculated by Livak and Schmittgen mathematical method [Methods (2001) 25(4):402-8.].
Results
Nine GLA alternative promoters were identified in the databases with overlapping sequences at 5-prime side of the GLA locus. EPD database showed 31 transcription starting sites (TSSs) along 150 bases (-99 to +50) from the main TSS at position 0. Three primers sets (G1, G2, and G3) were designed to cover several TSSs and were used in the GLA expression analysis by RT q-PCR. The expression data showed GLA expression differences over a wide range in kidney embryonic, epithelial and glomerular cells. For example, in the kidney embryonic cells, the GLA-G2 expression was decreased 12.5-fold, and GLA-G3 was increased 15.3-fold compared with GLA-G1 expression. Promoter efficiency depends upon regulatory sequences specific to transcription factors, mutations in the alternative promoters can alter the binding capacity of transcription factors.
Conclusion
RT q-PCR data showed the influence of alternative promoter type on the expression of the GLA gene in human kidney cells. Alternative promoters and their variants may serve as diagnostic biomarkers and potential therapeutic targets for Fabry disease.
Funding
- Commercial Support –