PUBLICATION

Identification of Cdca7 as a novel Notch transcriptional target involved in hematopoietic stem cell emergence

Authors
Guiu, J., Bergen, D.J., De Pater, E., Islam, A.B., Ayllón, V., Gama-Norton, L., Ruiz-Herguido, C., González, J., López-Bigas, N., Menendez, P., Dzierzak, E., Espinosa, L., Bigas, A.
ID
ZDB-PUB-141112-4
Date
2014
Source
The Journal of experimental medicine   211(12): 2411-23 (Journal)
Registered Authors
Bergen, Dylan
Keywords
none
MeSH Terms
  • Intercellular Signaling Peptides and Proteins/genetics
  • Intercellular Signaling Peptides and Proteins/metabolism
  • Membrane Proteins/genetics
  • Membrane Proteins/metabolism
  • Cell Line
  • Hematopoietic Stem Cells/cytology
  • Hematopoietic Stem Cells/metabolism*
  • Protein Binding
  • Mice, Knockout
  • Embryo, Mammalian/cytology
  • Embryo, Mammalian/embryology
  • Embryo, Mammalian/metabolism
  • Immunoglobulin J Recombination Signal Sequence-Binding Protein/genetics
  • Immunoglobulin J Recombination Signal Sequence-Binding Protein/metabolism
  • Nuclear Proteins/genetics*
  • Nuclear Proteins/metabolism
  • HEK293 Cells
  • Embryo, Nonmammalian/embryology
  • Embryo, Nonmammalian/metabolism
  • Zebrafish/embryology
  • Zebrafish/genetics
  • Animals
  • Aorta/embryology
  • Aorta/metabolism
  • Cell Differentiation/genetics
  • Gene Expression Regulation, Developmental
  • Transcription, Genetic*
  • In Situ Hybridization
  • Calcium-Binding Proteins/genetics
  • Calcium-Binding Proteins/metabolism
  • Reverse Transcriptase Polymerase Chain Reaction
  • Cells, Cultured
  • Receptor, Notch1/genetics*
  • Mice, Inbred C57BL
  • Humans
(all 35)
PubMed
25385755 Full text @ J. Exp. Med.
Abstract
Hematopoietic stem cell (HSC) specification occurs in the embryonic aorta and requires Notch activation; however, most of the Notch-regulated elements controlling de novo HSC generation are still unknown. Here, we identify putative direct Notch targets in the aorta-gonad-mesonephros (AGM) embryonic tissue by chromatin precipitation using antibodies against the Notch partner RBPj. By ChIP-on-chip analysis of the precipitated DNA, we identified 701 promoter regions that were candidates to be regulated by Notch in the AGM. One of the most enriched regions corresponded to the Cdca7 gene, which was subsequently confirmed to recruit the RBPj factor but also Notch1 in AGM cells. We found that during embryonic hematopoietic development, expression of Cdca7 is restricted to the hematopoietic clusters of the aorta, and it is strongly up-regulated in the hemogenic population during human embryonic stem cell hematopoietic differentiation in a Notch-dependent manner. Down-regulation of Cdca7 mRNA in cultured AGM cells significantly induces hematopoietic differentiation and loss of the progenitor population. Finally, using loss-of-function experiments in zebrafish, we demonstrate that CDCA7 contributes to HSC emergence in vivo during embryonic development. Thus, our study identifies Cdca7 as an evolutionary conserved Notch target involved in HSC emergence.
Genes / Markers
Figures
Figure Gallery (1 images)
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Expression
Phenotype
Mutations / Transgenics
Allele Construct Type Affected Genomic Region
hu5333TgTransgenic Insertion
    la2TgTransgenic Insertion
      1 - 2 of 2
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      Human Disease / Model
      No data available
      Sequence Targeting Reagents
      Target Reagent Reagent Type
      cdca7aMO1-cdca7aMRPHLNO
      cdca7aMO2-cdca7aMRPHLNO
      tp53MO4-tp53MRPHLNO
      1 - 3 of 3
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      Fish
      Antibodies
      No data available
      Orthology
      No data available
      Engineered Foreign Genes
      Marker Marker Type Name
      EGFPEFGEGFP
      RFPEFGRFP
      1 - 2 of 2
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      Mapping
      No data available