The Regulatory Factor ZFHX3 Modifies Circadian Function in SCN via an AT Motif-Driven Axis.
Bottom Line: Using RNA sequencing, we found minimal effects on core clock genes in Zfhx3(Sci/+) SCN, whereas the expression of neuropeptides critical for SCN intercellular signaling was significantly disturbed.Moreover, mutant ZFHX3 had a decreased ability to activate AT motifs in the promoters of these neuropeptide genes.Lentiviral transduction of SCN slices showed that the ZFHX3-mediated activation of AT motifs is circadian, with decreased amplitude and robustness of these oscillations in Zfhx3(Sci/+) SCN slices.
Affiliation: MRC Harwell, Harwell Science and Innovation Campus, Oxfordshire OX11 0RD, UK.Show MeSH
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Mentions: To confirm that ZFHX3 binds to the promoter of the key predicted target genes Avp and Vip in vivo, we performed quantitative ChIP on SCN tissue from Zfhx3+/+ animals (ZT3) using ZFHX3 antiserum. We found a significant increase in immunoprecipitated DNA upstream of the transcriptional start site (TSS) for both Avp and Vip compared to the Gapdh control promoter region, suggesting that ZFHX3 binds to both target gene promoters (Figures 6A and 6B). At the Vip promoter, this binding occurred in the vicinity of the AT motif to a far greater extent than in an adjacent upstream region. These data strongly suggest that ZFHX3 binds to Avp and Vip promoters in close proximity to AT consensus motifs.
Affiliation: MRC Harwell, Harwell Science and Innovation Campus, Oxfordshire OX11 0RD, UK.