Identification of an Enhancer Critical for the ephirn-A5 Gene Expression in the Posterior Region of the Mesencephalonopen access
- Authors
- Park, Eunjeong; Noh, Hyuna; Park, Soochul
- Issue Date
- Jun-2017
- Publisher
- KOREAN SOC MOLECULAR & CELLULAR BIOLOGY
- Keywords
- bHLH transcription factor; EphA; ephrin-A5; mesencephalon
- Citation
- MOLECULES AND CELLS, v.40, no.6, pp 426 - 433
- Pages
- 8
- Journal Title
- MOLECULES AND CELLS
- Volume
- 40
- Number
- 6
- Start Page
- 426
- End Page
- 433
- URI
- https://scholarworks.sookmyung.ac.kr/handle/2020.sw.sookmyung/8295
- DOI
- 10.14348/molcells.2017.0052
- ISSN
- 1016-8478
0219-1032
- Abstract
- Ephrin-A5 has been implicated in the regulation of brain morphogenesis and axon pathfinding. In this study, we used bacterial homologous recombination to express a LacZ reporter in various ephrin-A5 BAC clones to identify elements that regulate ephrin-A5 gene expression during mesencephalon development. We found that there is mesencephalonspecific enhancer activity localized to a specific + 25.0 kb to + 30.5 kb genomic region in the first intron of ephrin-A5. Further comparative genomic analysis indicated that two evolutionary conserved regions, ECR1 and ECR2, were present within this 5.5 kb region. Deletion of ECR1 from the enhancer resulted in disrupted mesencephalon-specific enhancer activity in transgenic embryos. We also found a consensus binding site for basic helix-loop-helix (bHLH) transcription factors (TFs) in a highly conserved region at the 3'-end of ECR1. We further demonstrated that specific deletion of the bHLH TF binding site abrogated the mesencephalon-specific enhancer activity in transgenic embryos. Finally, both electrophoretic mobility shift assay and luciferase-based transactivation assay revealed that the transcription factor Ascl1 bound the bHLH consensus binding site in the mesencephalonspecific ephrin-A5 enhancer in vitro. Together, these results suggest that the bHLH TF binding site in ECR1 is involved in the positive regulation of ephrin-A5 gene expression during the development of the mesencephalon.
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