Functional characterization of the ZEB2 regulatory landscape
| Autor: | Tommy Kaplan, Fania Shemuluvich, Ramon Y. Birnbaum, Reut Bar Yaacov, Reut Eshel, Einan Farhi |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
Central Nervous System Neurogenesis Hindbrain Biology Regulatory Sequences Nucleic Acid Cell Line 03 medical and health sciences Mice 0302 clinical medicine Notochord Genetics medicine Animals Humans Enhancer Molecular Biology Genetics (clinical) Conserved Sequence Sequence Deletion Zinc Finger E-box Binding Homeobox 2 Zinc finger Regulation of gene expression Neurons Base Sequence General Medicine Chromatin Cell biology 030104 developmental biology medicine.anatomical_structure Enhancer Elements Genetic Gene Expression Regulation Regulatory sequence Genetic Loci Organ Specificity Homeobox General Article 030217 neurology & neurosurgery Protein Binding |
| Zdroj: | Human Molecular Genetics |
| ISSN: | 1460-2083 0964-6906 |
| Popis: | Zinc finger E-box–binding homeobox 2 (ZEB2) is a key developmental regulator of the central nervous system (CNS). Although the transcriptional regulation of ZEB2 is essential for CNS development, the elements that regulate ZEB2 expression have yet to be identified. Here, we identified a proximal regulatory region of ZEB2 and characterized transcriptional enhancers during neuronal development. Using chromatin immunoprecipitation sequencing for active (H3K27ac) and repressed (H3K27me3) chromatin regions in human neuronal progenitors, combined with an in vivo zebrafish enhancer assay, we functionally characterized 18 candidate enhancers in the ZEB2 locus. Eight enhancers drove expression patterns that were specific to distinct mid/hindbrain regions (ZEB2#e3 and 5), trigeminal-like ganglia (ZEB2#e6 and 7), notochord (ZEB2#e2, 4 and 12) and whole brain (ZEB2#e14). We further dissected the minimal sequences that drive enhancer-specific activity in the mid/hindbrain and notochord. Using a reporter assay in human cells, we showed an increased activity of the minimal notochord enhancer ZEB2#e2 in response to AP-1 and DLX1/2 expressions, while repressed activity of this enhancer was seen in response to ZEB2 and TFAP2 expressions. We showed that Dlx1 but not Zeb2 and Tfap2 occupies Zeb2#e2 enhancer sequence in the mouse notochord at embryonic day 11.5. Using CRISPR/Cas9 genome editing, we deleted the ZEB2#e2 region, leading to reduction of ZEB2 expression in human cells. We thus characterized distal transcriptional enhancers and trans-acting elements that govern regulation of ZEB2 expression during neuronal development. These findings pave the path toward future analysis of the role of ZEB2 regulatory elements in neurodevelopmental disorders, such as Mowat–Wilson syndrome. |
| Databáze: | OpenAIRE |
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