Affiliations: [a] Department of Medical Genetics, Centre for Molecular Medicine and Therapeutics, BC Children’s Hospital, University of British Columbia, Vancouver, BC, Canada
| [b] Department of Clinical Neuroscience, Karolinska Institute, Stockholm, Sweden
Correspondence:
[*]
Correspondence to: Blair R. Leavitt, Center for Molecular Medicine and Therapeutics. 950 West 28th Avenue, Room 2020, Vancouver, BC V5Z 4H4, Canada. Tel.: +1 604 875 3801; Fax: +1 604 875 3840; E-mail: [email protected].
Note: [1] These authors contributed equally to this work.
Abstract: Background:Huntington’s disease is a late onset neurological disorder caused by a trinucleotide CAG repeat expansion mutation in the HTT gene encoding for the protein huntingtin. Despite considerable ongoing research, the wild-type function of huntingtin is not yet fully understood. Objective:To improve knowledge of HTT gene regulation at the transcriptional level and inform future studies aimed at uncovering the HTT gene’s normal function. Methods:The HTT gene region was functionally characterized through an in silico analysis 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 HTT promoter region. siRNA-mediated knockdown and ChIP-qPCR of STAT1, a TF identified from the in silico analysis, were used to validate the bioinformatics screen. Results: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 STAT1 resulted in an increase in HTT expression, and ChIP-qPCR detected enrichment of STAT1 binding at one of the predicted regions. These assays confirmed the utility of the bioinformatic analysis. Conclusions:Putative regulatory regions outside of the immediate HTT promoter region have been identified with specific protein-protein interactions. Future work will focus on in vitro and in vivo studies to examine the effect of modulating identified TFBSs and altering the levels of specific TFs of interest in regulating HTT gene expression.
