RFX5 regulates gene expression of the Pcdhα cluster
Received date: 2020-06-19
Revised date: 2020-07-08
Online published: 2020-07-29
Supported by
Supported by the National Natural Science Foundation of China Nos(91640118);Supported by the National Natural Science Foundation of China Nos(31630039)
Gene expression and three-dimensional (3D) genome organization are thought to be closely related. The protocadherin (Pcdh) clusters are central for neuronal self-avoidance in brain development and have been used as model genes to explore the role of 3D genome structures in gene regulation. Transcription factor RFX5 (regulatory factor x 5) is a member of the winged-helix subfamily of the helix-turn-helix superfamily proteins. The RFX5 protein contains four domains: oligomerization, DNA-binding, helical, and transactivation domains. RFX5 plays an important role in regulating expression of the major histocompatibility complex class II (MHC II) gene complex. Here we report a role of RFX5 in the regulation of clustered Pcdh genes. We first knocked out the RFX5 gene by using CRISPR/Cas9 DNA-fragment editing in HEC-1-B cell line. By RNA-seq experiments, we found that RFX5 deletion results in a significant increase of expression levels of the Pcdhα6, Pcdhα12 and Pcdhαc2 genes. By ChIP-nexus experiments, we found that RFX5 deletion leads to the increased binding of CTCF and RAD21 in the Pcdhα cluster. Finally, through quantitative high-resolution chromosome conformation capture copy (QHR-4C) experiments, we found that RFX5 deletion results in a significant increase of long-distance chromatin interactions between the HS5-1 enhancer and its target promoters in the Pcdhα cluster. In conclusion, RFX5 regulates gene expression of the Pcdhα cluster through higher-order chromatin structure.
Key words： RFX5; Pcdh; CTCF; cohesin; CRISPR/Cas9
Na Wang, Zhilian Jia, Qiang Wu . RFX5 regulates gene expression of the Pcdhα cluster[J]. Hereditas(Beijing), 2020 , 42(8) : 760 -774 . DOI: 10.16288/j.yczz.20-184
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