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

Cell Differentiation/genetics
Mice, Inbred C57BL
Immunoglobulin J Recombination Signal Sequence-Binding Protein/genetics
Immunoglobulin J Recombination Signal Sequence-Binding Protein/metabolism
Embryo, Nonmammalian/embryology
Embryo, Nonmammalian/metabolism
Cell Line
HEK293 Cells
Cells, Cultured
Protein Binding
Humans
Reverse Transcriptase Polymerase Chain Reaction
Zebrafish/embryology
Zebrafish/genetics
Embryo, Mammalian/cytology
Embryo, Mammalian/embryology
Embryo, Mammalian/metabolism
Transcription, Genetic*
Gene Expression Regulation, Developmental
Receptor, Notch1/genetics*
Animals
Nuclear Proteins/genetics*
Nuclear Proteins/metabolism
Hematopoietic Stem Cells/cytology
Hematopoietic Stem Cells/metabolism*
Intercellular Signaling Peptides and Proteins/genetics
Intercellular Signaling Peptides and Proteins/metabolism
In Situ Hybridization
Aorta/embryology
Aorta/metabolism
Membrane Proteins/genetics
Membrane Proteins/metabolism
Calcium-Binding Proteins/genetics
Calcium-Binding Proteins/metabolism
Mice, Knockout

(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

Show all Figures

Expression

Gene	Fish	Conditions	Stage	Qualifier	Anatomy	Assay	Figure
cdca7a	WT	control	Prim-15		ventral wall of dorsal aorta	ISH	Fig. 5
cdca7a	WT	chemical treatment: DAPT	Prim-15	Not Detected	ventral wall of dorsal aorta	ISH	Fig. 5
efnb2a	WT	standard conditions	Prim-15		dorsal aorta	ISH	Fig. 5
efnb2a	WT + MO2-cdca7a	standard conditions	Prim-15		dorsal aorta	ISH	Fig. 5
EGFP	hu5333Tg; la2Tg	standard conditions	Long-pec		hematopoietic stem cell	IFL	Fig. 5
EGFP	hu5333Tg; la2Tg	standard conditions	Long-pec		myeloid lineage restricted progenitor cell	IFL	Fig. 5
EGFP	hu5333Tg; la2Tg + MO2-cdca7a	standard conditions	Long-pec		hematopoietic stem cell	IFL	Fig. 5
EGFP	hu5333Tg; la2Tg + MO2-cdca7a	standard conditions	Long-pec		myeloid lineage restricted progenitor cell	IFL	Fig. 5
EGFP	la2Tg	standard conditions	Long-pec		hematopoietic stem cell	IFL	Fig. 5
EGFP	la2Tg	standard conditions	Long-pec		myeloid lineage restricted progenitor cell	IFL	Fig. 5

1 - 10 of 24

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Phenotype

Fish	Conditions	Stage	Phenotype	Figure
WT	chemical treatment: DAPT	Prim-15	Notch signaling pathway decreased occurrence, abnormal	Fig. 5
WT	chemical treatment: DAPT	Prim-15	ventral wall of dorsal aorta physical object quality, abnormal	Fig. 5
WT + MO2-cdca7a	standard conditions	Prim-15	artery development process quality, normal	Fig. 5
WT + MO2-cdca7a	standard conditions	Prim-15	hematopoietic stem cell differentiation decreased occurrence, abnormal	Fig. 5
hu5333Tg; la2Tg + MO2-cdca7a	standard conditions	Long-pec	hematopoietic stem cell differentiation decreased occurrence, abnormal	Fig. 5
hu5333Tg; la2Tg + MO2-cdca7a	standard conditions	Long-pec	whole organism has fewer parts of type hematopoietic stem cell, abnormal	Fig. 5

1 - 6 of 6

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Mutations / Transgenics

Allele	Construct	Type	Affected Genomic Region
hu5333Tg	Tg(-0.8flt1:RFP)	Transgenic Insertion
la2Tg	Tg(-6.0itga2b:EGFP)	Transgenic Insertion

Human Disease / Model

Sequence Targeting Reagents

Target	Reagent	Reagent Type
cdca7a	MO1-cdca7a	MRPHLNO
cdca7a	MO2-cdca7a	MRPHLNO
tp53	MO4-tp53	MRPHLNO

Fish

Fish
hu5333Tg
hu5333Tg; la2Tg
hu5333Tg; la2Tg + MO2-cdca7a
la2Tg
la2Tg + MO2-cdca7a
WT
WT + MO2-cdca7a

Antibodies

Orthology

Engineered Foreign Genes

Marker	Marker Type	Name
EGFP	EFG	EGFP
RFP	EFG	RFP

Mapping

Guiu et al., 2014 ZDB-PUB-141112-4 PMID:25385755

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

Guiu et al., 2014

ZDB-PUB-141112-4
PMID:25385755