PUBLICATION

Differential requirement for Gata1 DNA binding and transactivation between primitive and definitive stages of hematopoiesis in zebrafish

Authors

Belele, C.L., English, M.A., Chahal, J., Burnetti, A., Finckbeiner, S.M., Gibney, G., Kirby, M., Sood, R., and Liu, P.

ID

ZDB-PUB-091023-68

Date

2009

Source

Blood 114(25): 5162-5172 (Journal)

Registered Authors

English, Milton A., Liu, Pu Paul, Sood, Raman

Keywords

none

MeSH Terms

Female
Protein Structure, Tertiary
Mutation
Models, Molecular
Sequence Homology, Amino Acid
Gene Expression Regulation, Developmental
Zebrafish Proteins/chemistry
Zebrafish Proteins/genetics
Zebrafish Proteins/metabolism*
Hematopoiesis*
Male
Reverse Transcriptase Polymerase Chain Reaction
Amino Acid Sequence
Flow Cytometry
Molecular Sequence Data
Nucleic Acid Conformation
GATA1 Transcription Factor/chemistry
GATA1 Transcription Factor/genetics
GATA1 Transcription Factor/metabolism*
Animals
DNA/chemistry
DNA/metabolism*
Embryo, Nonmammalian/embryology
Embryo, Nonmammalian/metabolism
Zebrafish/embryology
Zebrafish/genetics
Zebrafish/metabolism*
Binding Sites/genetics
In Situ Hybridization
Protein Binding
Transcriptional Activation

(all 31)

PubMed

19843882 Full text @ Blood

Abstract

The transcription factor Gata1 is required for the development of erythrocytes and megakaryocytes. Previous studies with a complementation rescue approach showed that the zinc finger domains are required for both primitive and definitive hematopoiesis. Here we report a novel zebrafish gata1 mutant with an ENU-induced point mutation in the C-finger (gata1(T301K)). The Gata1 protein with this mutation bound to its DNA target sequence with reduced affinity and transactivated inefficiently in a reporter assay. gata1(T301K/T301K) fish had decreased number of erythrocytes during primitive hematopoiesis but normal adult hematopoiesis. We crossed the gata1(T301K/T301K) fish with those carrying the R339X mutation, also known as vlad tepes (vlt), which abolishes DNA binding and transactivation activities. As we reported before, gata1(vlt/vlt) embryos were "bloodless" and died around 11-15 days post fertilization (dpf). Interestingly, the gata1(T301K/vlt) fish had nearly complete block of primitive hematopoiesis, but they resumed hematopoiesis between 7-14 dpf and grew to phenotypically normal fish with normal adult hematopoiesis. Our findings suggest that the impact of Gata1 on hematopoiesis correlates with its DNA-binding ability and that primitive hematopoiesis is more sensitive to reduction in Gata1 function than definitive hematopoiesis.

Genes / Markers

Figures

Show all Figures

Expression

Phenotype

Fish	Conditions	Stage	Phenotype	Figure
gata1a^hg2/+; gata1a^m651/+	standard conditions	14-19 somites	nucleate erythrocyte maturation disrupted, abnormal	Fig. 3
gata1a^hg2/+; gata1a^m651/+	standard conditions	14-19 somites	primitive hemopoiesis disrupted, abnormal	Fig. 3
gata1a^hg2/+; gata1a^m651/+	standard conditions	Prim-5	macrophage differentiation increased occurrence, abnormal	Fig. 5
gata1a^hg2/+; gata1a^m651/+	standard conditions	Prim-5	monocyte differentiation increased occurrence, abnormal	Fig. 5
gata1a^hg2/+; gata1a^m651/+	standard conditions	Prim-5	neutrophil differentiation occurrence, normal	Fig. 5
gata1a^hg2/+; gata1a^m651/+	standard conditions	Prim-5	primitive hemopoiesis disrupted, abnormal	Fig. 5
gata1a^hg2/+; gata1a^m651/+	standard conditions	Prim-5 to Day 4	nucleate erythrocyte present, abnormal	Fig. 4
gata1a^hg2/+; gata1a^m651/+	standard conditions	Long-pec	nucleate erythrocyte absent, abnormal	Fig. 2
gata1a^hg2/+; gata1a^m651/+	standard conditions	Long-pec	primitive hemopoiesis disrupted, abnormal	Fig. 2
gata1a^hg2/+; gata1a^m651/+	standard conditions	Days 7-13 to Days 14-20	nucleate erythrocyte absent, abnormal	Fig. 4

1 - 10 of 24

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Mutations / Transgenics

Allele	Construct	Type	Affected Genomic Region
hg2		Point Mutation	gata1a
hg3		Point Mutation	gata1a
la781Tg	Tg(gata1a:GFP)	Transgenic Insertion
m651		Point Mutation	gata1a

Human Disease / Model

Sequence Targeting Reagents

Target	Reagent	Reagent Type
gata2a	MO1-gata2a	MRPHLNO

Fish

Fish
gata1a^hg2/hg2
gata1a^hg2/hg2; la781Tg
gata1a^hg2/+
gata1a^hg2/+; gata1a^m651/+
gata1a^hg2/+; gata1a^m651/+; la781Tg
gata1a^hg3/hg3
gata1a^m651/m651
gata1a^m651/m651 + MO1-gata2a
WT

Antibodies

Orthology

Engineered Foreign Genes

Marker	Marker Type	Name
GFP	EFG	GFP

Mapping

Belele et al., 2009 ZDB-PUB-091023-68 PMID:19843882

Differential requirement for Gata1 DNA binding and transactivation between primitive and definitive stages of hematopoiesis in zebrafish

Belele et al., 2009

ZDB-PUB-091023-68
PMID:19843882