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PMID:19843882

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Citation

Belele, CL, English, MA, Chahal, J, Burnetti, A, Finckbeiner, SM, Gibney, G, Kirby, M, Sood, R and Liu, PP (2009) Differential requirement for Gata1 DNA binding and transactivation between primitive and definitive stages of hematopoiesis in zebrafish. Blood 114:5162-72

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 N-ethyl-N-nitrosourea-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 a 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 previously, gata1(vlt/vlt) embryos were "bloodless" and died approximately 11 to 15 days after fertilization. Interestingly, the gata1(T301K/vlt) fish had nearly a complete block of primitive hematopoiesis, but they resumed hematopoiesis between 7 and 14 days after fertilization 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.

Links

PubMed PMC2792212 Online version:10.1182/blood-2009-05-224709

Keywords

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

Significance

Annotations

Gene product Qualifier GO Term Evidence Code with/from Aspect Extension Notes Status

DANRE:A0A0R4IKY6

involved_in

GO:0060215: primitive hemopoiesis

ECO:0000316: genetic interaction evidence used in manual assertion

ZFIN:ZDB-GENO-070810-1,ZFIN:ZDB-MRPHLNO-050208-12

P

Seeded From UniProt

complete

DANRE:A0A2R9YJP8

involved_in

GO:0060215: primitive hemopoiesis

ECO:0000316: genetic interaction evidence used in manual assertion

ZFIN:ZDB-GENO-070810-1,ZFIN:ZDB-MRPHLNO-050208-12

P

Seeded From UniProt

complete

DANRE:O42355

involved_in

GO:0060215: primitive hemopoiesis

ECO:0000316: genetic interaction evidence used in manual assertion

ZFIN:ZDB-GENO-070810-1,ZFIN:ZDB-MRPHLNO-050208-12

P

Seeded From UniProt

complete

DANRE:Q05AJ3

involved_in

GO:0030225: macrophage differentiation

ECO:0000315: mutant phenotype evidence used in manual assertion

ZFIN:ZDB-GENO-100722-3

P

Seeded From UniProt

complete

DANRE:Q05AJ3

involved_in

GO:0030225: macrophage differentiation

ECO:0000315: mutant phenotype evidence used in manual assertion

ZFIN:ZDB-GENO-070810-1

P

Seeded From UniProt

complete

DANRE:Q05AJ3

involved_in

GO:0030224: monocyte differentiation

ECO:0000315: mutant phenotype evidence used in manual assertion

ZFIN:ZDB-GENO-100722-3

P

Seeded From UniProt

complete

DANRE:Q05AJ3

involved_in

GO:0030224: monocyte differentiation

ECO:0000315: mutant phenotype evidence used in manual assertion

ZFIN:ZDB-GENO-070810-1

P

Seeded From UniProt

complete

DANRE:Q05AJ3

enables

GO:0003700: DNA-binding transcription factor activity

ECO:0000314: direct assay evidence used in manual assertion

F

Seeded From UniProt

complete

DANRE:Q05AJ3

enables

GO:0043565: sequence-specific DNA binding

ECO:0000314: direct assay evidence used in manual assertion

F

Seeded From UniProt

complete

DANRE:Q05AJ3

NOT|involved_in

GO:0060216: definitive hemopoiesis

ECO:0000315: mutant phenotype evidence used in manual assertion

ZFIN:ZDB-GENO-100722-3

P

Seeded From UniProt

complete

DANRE:Q05AJ3

NOT|involved_in

GO:0060216: definitive hemopoiesis

ECO:0000315: mutant phenotype evidence used in manual assertion

ZFIN:ZDB-GENO-070810-1

P

Seeded From UniProt

complete

DANRE:Q05AJ3

involved_in

GO:0060215: primitive hemopoiesis

ECO:0000315: mutant phenotype evidence used in manual assertion

ZFIN:ZDB-GENO-100722-3

P

Seeded From UniProt

complete

DANRE:Q05AJ3

involved_in

GO:0060215: primitive hemopoiesis

ECO:0000315: mutant phenotype evidence used in manual assertion

ZFIN:ZDB-GENO-070810-1

P

Seeded From UniProt

complete

DANRE:Q05AJ3

involved_in

GO:0060215: primitive hemopoiesis

ECO:0000316: genetic interaction evidence used in manual assertion

ZFIN:ZDB-GENO-070810-1,ZFIN:ZDB-MRPHLNO-050208-12

P

Seeded From UniProt

complete

DANRE:Q7T3G1

involved_in

GO:0060215: primitive hemopoiesis

ECO:0000316: genetic interaction evidence used in manual assertion

ZFIN:ZDB-GENO-070810-1,ZFIN:ZDB-MRPHLNO-050208-12

P

Seeded From UniProt

complete

DANRE:Q90410

involved_in

GO:0030225: macrophage differentiation

ECO:0000315: mutant phenotype evidence used in manual assertion

ZFIN:ZDB-GENO-100722-3

P

Seeded From UniProt

complete

DANRE:Q90410

involved_in

GO:0030225: macrophage differentiation

ECO:0000315: mutant phenotype evidence used in manual assertion

ZFIN:ZDB-GENO-070810-1

P

Seeded From UniProt

complete

DANRE:Q90410

involved_in

GO:0030224: monocyte differentiation

ECO:0000315: mutant phenotype evidence used in manual assertion

ZFIN:ZDB-GENO-100722-3

P

Seeded From UniProt

complete

DANRE:Q90410

involved_in

GO:0030224: monocyte differentiation

ECO:0000315: mutant phenotype evidence used in manual assertion

ZFIN:ZDB-GENO-070810-1

P

Seeded From UniProt

complete

DANRE:Q90410

enables

GO:0003700: DNA-binding transcription factor activity

ECO:0000314: direct assay evidence used in manual assertion

F

Seeded From UniProt

complete

DANRE:Q90410

enables

GO:0043565: sequence-specific DNA binding

ECO:0000314: direct assay evidence used in manual assertion

F

Seeded From UniProt

complete

DANRE:Q90410

NOT|involved_in

GO:0060216: definitive hemopoiesis

ECO:0000315: mutant phenotype evidence used in manual assertion

ZFIN:ZDB-GENO-100722-3

P

Seeded From UniProt

complete

DANRE:Q90410

NOT|involved_in

GO:0060216: definitive hemopoiesis

ECO:0000315: mutant phenotype evidence used in manual assertion

ZFIN:ZDB-GENO-070810-1

P

Seeded From UniProt

complete

DANRE:Q90410

involved_in

GO:0060215: primitive hemopoiesis

ECO:0000315: mutant phenotype evidence used in manual assertion

ZFIN:ZDB-GENO-100722-3

P

Seeded From UniProt

complete

DANRE:Q90410

involved_in

GO:0060215: primitive hemopoiesis

ECO:0000315: mutant phenotype evidence used in manual assertion

ZFIN:ZDB-GENO-070810-1

P

Seeded From UniProt

complete

DANRE:Q90410

involved_in

GO:0060215: primitive hemopoiesis

ECO:0000316: genetic interaction evidence used in manual assertion

ZFIN:ZDB-GENO-070810-1,ZFIN:ZDB-MRPHLNO-050208-12

P

Seeded From UniProt

complete

DANRE:Q90411

involved_in

GO:0060215: primitive hemopoiesis

ECO:0000316: genetic interaction evidence used in manual assertion

ZFIN:ZDB-GENO-070810-1,ZFIN:ZDB-MRPHLNO-050208-12

P

Seeded From UniProt

complete


See also

References

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