| Autor: |
De Souza, Rebecca A.G., Kosior, Natalia, Thomson, Sarah B., Mathelier, Anthony, Zhang, Allen W., Bečanović, Kristina, Wasserman, Wyeth W., Leavitt, Blair R. |
| Zdroj: |
Journal of Huntington's Disease; July 2018, Vol. 7 Issue: 3 p223-237, 15p |
| Abstrakt: |
Huntington’s disease is a late onset neurological disorder caused by a trinucleotide CAG repeat expansion mutation in the HTTgene encoding for the protein huntingtin. Despite considerable ongoing research, the wild-type function of huntingtin is not yet fully understood. To improve knowledge of HTTgene regulation at the transcriptional level and inform future studies aimed at uncovering the HTTgene’s normal function. The HTTgene region was functionally characterized through an in silicoanalysis using publicly available data sets. ChIP-seq data sets and the online STRING database were used to identify putative transcription factor binding sites (TFBSs) and protein-protein interactions within the HTTpromoter region. siRNA-mediated knockdown and ChIP-qPCR of STAT1, a TF identified from the in silicoanalysis, were used to validate the bioinformatics screen. 16 regions containing potential regulatory genomic markers were identified. TFBSs for 59 transcription factors (TFs) were detected in one or more of the 16 candidate regions. Using these TFs, 15 clusters of protein-protein interactions were identified using STRING. siRNA-mediated knockdown of STAT1resulted in an increase in HTTexpression, and ChIP-qPCR detected enrichment of STAT1 binding at one of the predicted regions. These assays confirmed the utility of the bioinformatic analysis. Putative regulatory regions outside of the immediate HTTpromoter region have been identified with specific protein-protein interactions. Future work will focus on in vitroand in vivostudies to examine the effect of modulating identified TFBSs and altering the levels of specific TFs of interest in regulating HTTgene expression. |
| Databáze: |
Supplemental Index |
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