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ZIRC
ZFIN ID: ZDB-PUB-091023-68
Differential requirement for Gata1 DNA binding and transactivation between primitive and definitive stages of hematopoiesis in zebrafish
Belele, C.L., English, M.A., Chahal, J., Burnetti, A., Finckbeiner, S.M., Gibney, G., Kirby, M., Sood, R., and Liu, P.
Date: 2009
Source: Blood 114(25): 5162-5172 (Journal)
Registered Authors: English, Milton A., Liu, Pu Paul, Sood, Raman
Keywords: none
MeSH Terms:
  • 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*
PubMed: 19843882 Full text @ Blood
FIGURES
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.
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