Identification of potential target genes for RFX4_v3, a transcription factor critical for brain development
Version of Record online: 26 JUN 2006
Journal of Neurochemistry
Volume 98, Issue 3, pages 860–875, August 2006
How to Cite
Zhang, D., Stumpo, D. J., Graves, J. P., DeGraff, L. M., Grissom, S. F., Collins, J. B., Li, L., Zeldin, D. C. and Blackshear, P. J. (2006), Identification of potential target genes for RFX4_v3, a transcription factor critical for brain development. Journal of Neurochemistry, 98: 860–875. doi: 10.1111/j.1471-4159.2006.03930.x
- Issue online: 26 JUN 2006
- Version of Record online: 26 JUN 2006
- Received December 19, 2005; revised manuscript received March 22, 2006; accepted March 24, 2006.
- CX3C ligand 1;
- regulatory factor X4 variant transcript 3.
Regulatory factor X4 variant transcript 3 (Rfx4_v3) gene disruption in mice demonstrated that interruption of a single allele (heterozygous, +/–) prevented formation of the subcommissural organ, resulting in congenital hydrocephalus, while interruption of two alleles (homozygous, –/–) caused fatal failure of dorsal midline brain structure formation. To identify potential target genes for RFX4_v3, we used microarray analysis to identify differentially expressed genes in Rfx4_v3-deficient mouse brains at embryonic day 10.5, before gross structural changes were apparent. Of 109 differentially expressed transcripts, 24 were chosen for validation and 22 were confirmed by real-time PCR. Many validated genes encoded critical proteins involved in brain morphogenesis, such as the signaling components in the Wnt, bone morphogenetic protein (BMP) and retinoic acid (RA) pathways. Cx3cl1, a CX3C-type chemokine gene that is highly expressed in brain, was down-regulated in the Rfx4_v3-null mice. Both human and mouse Cx3cl1 proximal promoters contained highly conserved X-boxes, known cis-acting elements for RFX protein binding. Using the Cx3cl1 promoter as an example of a target gene, we demonstrated direct binding of RFX4_v3 to the Cx3cl1 promoter, and trans-acting activity of RFX4_v3 protein to stimulate gene expression. These data suggest that RFX4_v3 may act upstream of critical signaling pathways in the process of brain development.