Regulation Of Erythroid Gene Expression

National Institute Of Diabetes And Digestive And Kidney Diseases

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Abstract

PU.1 is a transcription factor that interacts with the GATA-1 DNA binding domain, and there is reciprocal inhibition between the two proteins that determines cell fate within the hematopoietic lineages. The PU.1 transactivation (TAD) and DNA binding domains are reported to be involved in the interaction and we have noted that the PU.1 TAD shares structural homology with the TAD of p53. In collaboration with the laboratory of Jim Omichinski we have shown that the p53 TAD also interacts in vitro and in vivo with the GATA-1 DNA binding domain. The linker and C-finger of GATA-1 are required for the interaction. The proteins reciprocally inhibit the transactivation activity of one another in an erythroid precursor cell line, 6C2. GATA-1 may be required to prevent p53 induction during the nuclear condensation and enucleation that precedes erythrocyte formation or during the polyploidization of megakaryocytes. In collaboration with Masi Yamamoto we are testing mutants of GATA-1 that do not interact with p53 in rescue assays in GATA1.05 cells and mice to determine the role of this interaction during hematopoiesis. Another project involves anemias associated with the EKLF transcription factor. Mutations in the EKLF DNA binding domain occur in people with anemias. An amino acid in the second zinc finger of EKLF is mutated in patients with CBA and in the Nan (Neonatal anemia) mouse. In collaboration with Jim Bieker we are trying to determine if the anemic phenotype is based on different DNA binding affinities of Nan-EKLF relative to wild-type for some EKLF target genes.