| TF experiment description: |
We hypothesized that these SNP variants could disrupt transcription factor binding, altering enhancer activity for each allele. At rs6758183, the disease associated A allele increased the strength of a SMAD motif, and decreased the strength of an ETS motif (Figure 7D). Using ChIP, we found that EHF bound to the WT allele of rs6758183, suggesting disruption of EHF binding by the SNP could result in the observed differences in enhancer activity (Figure 7F). Intriguingly, the WT version of the SNP showed less enhancer activity than the mutant, suggesting EHF binding acts to repress the enhancer region. When this binding is lost, the enhancer becomes overactive, a change that could result in growth imbalances that cause alterations in the curvature and refractive power of the cornea. Consistent with this possibility, in previously published siRNA data for EHF in corneal epithelial cells, we found that knockdown of EHF caused a small but significant increase in IRS1 gene expression (Figure 7G) (18). Together, these data suggest EHF regulates IRS1 through binding to the rs6758183 enhancer and that disruption of the EHF motif causes reduced affinity of EHF for this site, resulting in aberrant IRS1 expression during corneal development. ;We hypothesized that these SNP variants could disrupt transcription factor binding, altering enhancer activity for each allele. At rs6758183, the disease associated A allele increased the strength of a SMAD motif, and decreased the strength of an ETS motif (Figure 7D). Using ChIP, we found that EHF bound to the WT allele of rs6758183, suggesting disruption of EHF binding by the SNP could result in the observed differences in enhancer activity (Figure 7F). Intriguingly, the WT version of the SNP showed less enhancer activity than the mutant, suggesting EHF binding acts to repress the enhancer region. When this binding is lost, the enhancer becomes overactive, a change that could result in growth imbalances that cause alterations in the curvature and refractive power of the cornea. Consistent with this possibility, in previously published siRNA data for EHF in corneal epithelial cells, we found that knockdown of EHF caused a small but significant increase in IRS1 gene expression (Figure 7G) (18). Together, these data suggest EHF regulates IRS1 through binding to the rs6758183 enhancer and that disruption of the EHF motif causes reduced affinity of EHF for this site, resulting in aberrant IRS1 expression during corneal development. ;We hypothesized that these SNP variants could disrupt transcription factor binding, altering enhancer activity for each allele. At rs6758183, the disease associated A allele increased the strength of a SMAD motif, and decreased the strength of an ETS motif (Figure 7D). Using ChIP, we found that EHF bound to the WT allele of rs6758183, suggesting disruption of EHF binding by the SNP could result in the observed differences in enhancer activity (Figure 7F). Intriguingly, the WT version of the SNP showed less enhancer activity than the mutant, suggesting EHF binding acts to repress the enhancer region. When this binding is lost, the enhancer becomes overactive, a change that could result in growth imbalances that cause alterations in the curvature and refractive power of the cornea. Consistent with this possibility, in previously published siRNA data for EHF in corneal epithelial cells, we found that knockdown of EHF caused a small but significant increase in IRS1 gene expression (Figure 7G) (18). Together, these data suggest EHF regulates IRS1 through binding to the rs6758183 enhancer and that disruption of the EHF motif causes reduced affinity of EHF for this site, resulting in aberrant IRS1 expression during corneal development. ;We hypothesized that these SNP variants could disrupt transcription factor binding, altering enhancer activity for each allele. At rs6758183, the disease associated A allele increased the strength of a SMAD motif, and decreased the strength of an ETS motif (Figure 7D). Using ChIP, we found that EHF bound to the WT allele of rs6758183, suggesting disruption of EHF binding by the SNP could result in the observed differences in enhancer activity (Figure 7F). Intriguingly, the WT version of the SNP showed less enhancer activity than the mutant, suggesting EHF binding acts to repress the enhancer region. When this binding is lost, the enhancer becomes overactive, a change that could result in growth imbalances that cause alterations in the curvature and refractive power of the cornea. Consistent with this possibility, in previously published siRNA data for EHF in corneal epithelial cells, we found that knockdown of EHF caused a small but significant increase in IRS1 gene expression (Figure 7G) (18). Together, these data suggest EHF regulates IRS1 through binding to the rs6758183 enhancer and that disruption of the EHF motif causes reduced affinity of EHF for this site, resulting in aberrant IRS1 expression during corneal development. |
