Sox2 Transcriptionally Regulates Pqbp1, an Intellectual Disability-Microcephaly Causative Gene, in Neural Stem Progenitor Cells

Abstract

PQBP1 is a nuclear-cytoplasmic shuttling protein that is engaged in RNA metabolism and transcription. In mouse embryonic brain, our previous in situ hybridization study revealed that PQBP1 mRNA was dominantly expressed in the periventricular zone region where neural stem progenitor cells (NSPCs) are located. Because the expression patterns in NSPCs are related to the symptoms of intellectual disability and microcephaly in PQBP1 gene-mutated patients, we investigated the transcriptional regulation of PQBP1 by NSPC-specific transcription factors. We selected 132 genome sequences that matched the consensus sequence for the binding of Sox2 and POU transcription factors upstream and downstream of the mouse PQBP1 gene. We then screened the binding affinity of these sequences to Sox2-Pax6 or Sox2-Brn2 with gel mobility shift assays and found 18 genome sequences that interacted with the NSPC-specific transcription factors. Some of these sequences had cis-regulatory activities in Luciferase assays and in utero electroporation into NSPCs. Furthermore we found decreased levels of expression of PQBP1 protein in NSPCs of heterozygous Sox2-knockout mice in vivo by immunohistochemistry and western blot analysis. Collectively, these results indicated that Sox2 regulated the transcription of PQBP1 in NSPCs.

Topographical transcriptome mapping of the mouse medial ganglionic eminence by spatially resolved RNA-seq

“…In each case, in situ hybridization was combined with immunohistochemistry for Gap43, which defines cluster 4/5 (Figure 4 ). Sox 2 [ 27 - 29 ] and COUP transcription factor 1 ( Couptf1 ), also known as Nr2f2 , [ 26 , 30 ] were chosen as markers for cluster 1, and in situ hybridization for both genes showed their expression restricted to the ventricular zone, as expected (Figure 4 A-H). For cluster 2, in situ hybridization for Doublecortin like kinase 2 ( Dclk2 ) and Sox11 [ 31 , 32 ] delineated a narrow region immediately ventral to the ventricular zone, matching the expected location of this cluster (Figure 4 I-L).…”

Topographical transcriptome mapping of the mouse medial ganglionic eminence by spatially-resolved RNA-seq

“…In each case, in situ hybridization was combined with immunohistochemistry for Gap43, which defines cluster 4/5 ( Figure 4). Sox 2 [27][28][29] and COUP transcription factor 1 (Couptf1), also known as Nr2f2, [26,30] were chosen as markers for cluster 1, and in situ hybridization for both genes showed their expression restricted to the ventricular zone, as expected ( Figure 4A-H). For cluster 2, in situ hybridization for Doublecortin like kinase 2 (Dclk2) and Sox11 [31,32] delineated a narrow region immediately ventral to the ventricular zone, matching the expected location of this cluster ( Figure 4I-L).…”

In utero gene therapy rescues microcephaly caused by Pqbp1-hypofunction in neural stem progenitor cells

“…Pqbp1 is highly expressed in NSPCs (Sox2-positive AP cells and RC2-positive radial glia in the VZ and SVZ) and Sox2 transcriptionally regulates Pqbp1 in NSPCs. 27 Pqbp1 is also expressed in differentiated neurons at a lower level. 5 , 27 , 28 We therefore generated two types of conditional KO (cKO) mice using nestin-Cre and synapsin-1-Cre ( Figures 1d–g , and Supplementary Figure 1A ).…”

“… 27 Pqbp1 is also expressed in differentiated neurons at a lower level. 5 , 27 , 28 We therefore generated two types of conditional KO (cKO) mice using nestin-Cre and synapsin-1-Cre ( Figures 1d–g , and Supplementary Figure 1A ). As expected, the expression level of the Pqbp1 protein was remarkably decreased in the total brain tissue of nestin-Cre cKO mice, although a faint band was detected due to the non-neural tissues in the brain ( Supplementary Figure 1B ).…”