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

Autor: Leonor Gama-Norton, Nuria Lopez-Bigas, Emma de Pater, Verónica Ayllón, Abul B. M. M. K. Islam, Lluis Espinosa, Cristina Ruiz-Herguido, Elaine Dzierzak, Anna Bigas, Jessica González, Pablo Menendez, Jordi Guiu, Dylan J. M. Bergen
Přispěvatelé: Cell biology
Rok vydání: 2014
Předmět:
Embryo
Nonmammalian
Transcription
Genetic
Cellular differentiation
Immunology
Population
Article
Cell Line
medicine
Animals
Humans
Immunology and Allergy
Serrate-Jagged Proteins
Receptor
Notch1
education
Zebrafish
Aorta
Cells
Cultured
In Situ Hybridization
Mice
Knockout
education.field_of_study
biology
Reverse Transcriptase Polymerase Chain Reaction
RBPJ
Calcium-Binding Proteins
HEK 293 cells
Gene Expression Regulation
Developmental
Membrane Proteins
Nuclear Proteins
Hematopoietic stem cell
Cell Differentiation
Cell Biology
Embryonic Tissue
Embryo
Mammalian
Hematopoietic Stem Cells
biology.organism_classification
Embryonic stem cell
Molecular biology
Cultius cel·lulars
Cell biology
Mice
Inbred C57BL
HEK293 Cells
medicine.anatomical_structure
Immunoglobulin J Recombination Signal Sequence-Binding Protein
Intercellular Signaling Peptides and Proteins
Peixos zebra (Animals de laboratori)
Protein Binding
Zdroj: Journal of Experimental Medicine, 211(12), 2411-2423. Rockefeller University Press
The Journal of Experimental Medicine
Recercat. Dipósit de la Recerca de Catalunya
instname
Guiu, J, Bergen, D J M, De Pater, E, Islam, A B M M K, Ayllón, V, Gama-Norton, L, Ruiz-Herguido, C, González, J, López-Bigas, N, Menendez, P, Dzierzak, E, Espinosa, L & Bigas, A 2014, ' Identification of Cdca7 as a novel Notch transcriptional target involved in hematopoietic stem cell emergence ', Journal of Experimental Medicine, vol. 211, no. 12, pp. 2411-23 . https://doi.org/10.1084/jem.20131857
Europe PubMed Central
ISSN: 0022-1007
DOI: 10.1084/jem.20131857
Popis: Guiu et al. use ChIP-on-chip analysis for the Notch partner RBPj, using embryonic tissue from the aorta-gonad-mesonephros region to identify potential novel Notch target genes involved in HSC emergence. They show that c-MYC–responsive gene Cdca7 is expressed in different HSC and progenitor subpopulations and that CDCA7 is important for maintaining the undifferentiated phenotype. Cdca7 acts downstream of Notch in HSCs in zebrafish, mouse, and human, indicating a highly conserved Notch/RBPj/Cdca7 axis in hematopoietic development.
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.
Databáze: OpenAIRE
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