PUBLICATION

Intronic Flk1 Enhancer Directs Arterial-Specific Expression via RBPJ-Mediated Venous Repression

Authors
Becker, P.W., Sacilotto, N., Nornes, S., Neal, A., Thomas, M., Liu, K., Preece, C., Ratnayaka, I., Davies, B., Bou-Gharios, G., De Val, S.
ID
ZDB-PUB-160417-9
Date
2016
Source
Arteriosclerosis, Thrombosis, and Vascular Biology   36(6): 1209-19 (Journal)
Registered Authors
Becker, Philipp, De Val, Sarah, Neal, Alice, Nornes, Svanhild, Sacilotto, Natalia
Keywords
artery, endothelial cells, mice, veins, zebrafish
MeSH Terms
  • Genes, Reporter
  • Immunoglobulin J Recombination Signal Sequence-Binding Protein/genetics
  • Immunoglobulin J Recombination Signal Sequence-Binding Protein/metabolism*
  • Arteries/embryology
  • Arteries/metabolism*
  • Animals
  • Signal Transduction
  • Receptors, Notch/metabolism
  • Mutation
  • Zebrafish Proteins/genetics
  • Zebrafish Proteins/metabolism*
  • SOX Transcription Factors/metabolism
  • Vascular Endothelial Growth Factor A/metabolism
  • Binding Sites
  • Vascular Endothelial Growth Factor Receptor-2/genetics
  • Vascular Endothelial Growth Factor Receptor-2/metabolism*
  • Proto-Oncogene Proteins c-ets/metabolism
  • GATA Transcription Factors/metabolism
  • Gene Silencing
  • Endothelial Cells/metabolism*
  • Mutagenesis, Site-Directed
  • Neovascularization, Physiologic*
  • Enhancer Elements, Genetic
  • Introns
  • Gene Expression Regulation, Developmental
  • Veins/embryology
  • Veins/metabolism*
  • Mice, Transgenic
  • Zebrafish/embryology
  • Zebrafish/genetics
  • Zebrafish/metabolism*
(all 31)
PubMed
27079877 Full text @ Arterio., Thromb., and Vas. Bio.
Abstract
The vascular endothelial growth factor receptor Flk1 is essential for vascular development, but the signaling and transcriptional pathways by which its expression is regulated in endothelial cells remain unclear. Although previous studies have identified 2 Flk1 regulatory enhancers, these are dispensable for Flk1 expression, indicating that additional enhancers contribute to Flk1 regulation in endothelial cells. In the present study, we sought to identify Flk1 enhancers contributing to expression in endothelial cells.
A region of the 10th intron of the Flk1 gene (Flk1in10) was identified as a putative enhancer and tested in mouse and zebrafish transgenic models. This region robustly directed reporter gene expression in arterial endothelial cells. Using a combination of targeted mutagenesis of transcription factor-binding sites and gene silencing of transcription factors, we found that Gata and Ets factors are required for Flk1in10 enhancer activity in all endothelial cells. Furthermore, we showed that activity of the Flk1in10 enhancer is restricted to arteries through repression of gene expression in venous endothelial cells by the Notch pathway transcriptional regulator Rbpj.
This study demonstrates a novel mechanism of arterial-venous identity acquisition, indicates a direct link between the Notch and vascular endothelial growth factor signaling pathways, and illustrates how cis-regulatory diversity permits differential expression outcomes from a limited repertoire of transcriptional regulators.
Genes / Markers
Figures
Figure Gallery (6 images)
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Expression
Phenotype
No data available
Mutations / Transgenics
Allele Construct Type Affected Genomic Region
lcr2TgTransgenic Insertion
    s896TgTransgenic Insertion
      1 - 2 of 2
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      Human Disease / Model
      No data available
      Sequence Targeting Reagents
      Target Reagent Reagent Type
      gata1aMO1-gata1aMRPHLNO
      gata2aMO1-gata2aMRPHLNO
      rbpjaMO4-rbpja,rbpjbMRPHLNO
      rbpjbMO4-rbpja,rbpjbMRPHLNO
      sox7MO5-sox7MRPHLNO
      sox18MO5-sox18MRPHLNO
      tnnt2aMO1-tnnt2aMRPHLNO
      1 - 7 of 7
      Show
      Fish
      No data available
      Antibodies
      No data available
      Orthology
      No data available
      Engineered Foreign Genes
      Marker Marker Type Name
      GFPEFGGFP
      mCherryEFGmCherry
      1 - 2 of 2
      Show
      Mapping
      No data available