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.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.In conclusion, by cloning Zfhx3(Sci), we have uncovered a circadian transcriptional axis that determines the period and robustness of behavioral and SCN molecular rhythms.
Affiliation: MRC Harwell, Harwell Science and Innovation Campus, Oxfordshire OX11 0RD, UK.Show MeSH
Mentions: The limited work investigating this transcription factor to date has revealed a number of its gene targets (Berry et al., 2001; Mori et al., 2007; Qi et al., 2008; Yasuda et al., 1994). Using a phylogenetic shadowing-based approach (Bailey and Elkan, 1994), we constructed a motif-binding model for ZFHX3 based on the characterized binding motifs in these promoters (see Supplemental Experimental Procedures for more details) (Figure 3A). We then investigated whether this consensus AT motif, as well as the canonical circadian E-box, D-box, and RRE, were present in sequences upstream of the genes differentially regulated by ZFHX3 (Pscan score > 0.88). We found that the AT motif was present in promoter sequences from 39% of the differentially expressed genes, while the circadian motifs E-box, D-box, and RRE were absent (Figure 3B).
Affiliation: MRC Harwell, Harwell Science and Innovation Campus, Oxfordshire OX11 0RD, UK.