Regulation Of Erythroid Gene Expression

红系基因表达的调控

基本信息

批准号：
8349637
负责人：
Gary Felsenfeld
金额：
$ 27.67万
依托单位：
NATIONAL INSTITUTE OF DIABETES AND DIGESTIVE AND KIDNEY DISEASES
依托单位国家：
美国
项目类别：
财政年份：
资助国家：
美国
起止时间：
至
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/8349637
关键词：
Acetylation Affect Affinity Amino Acids Anemia Binding Binding Sites Biochemical Biological Assay C-terminal Cell Line Cells Characteristics Chromatin Chromatin Structure Collaborations DNA DNA Binding DNA Binding Domain Development EP300 gene Embryo Erythrocytes Erythroid Erythroid Progenitor Cells Erythropoiesis Family Fingers Fogs Gene Expression Gene Targeting Hematopoiesis Hematopoietic Human In Vitro Laboratories Lysine Megakaryocytes Modification Mus Mutate Mutation N-terminal Neonatal Anemia Nuclear Patients Peptides Phenotype Phosphorylation Physical condensation Point Mutation Proteins Regulation Relative (related person)Reporting Role Site Specific qualifier value Specificity Structure Tars Testing Thalassemia Transactivation Transcription Repressor/Corepressor Ubiquitination Zinc Fingers arm base cofactor eosinophil erythroid Kruppel-like factor human GATA1 protein in vivo interest leukemia mast cell mutant preference prevent protein protein interaction research study transcription factor

项目摘要

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.

PU.1是与GATA-1 DNA结合相互作用的转录因子结构域，两种蛋白质之间存在互惠抑制，这些蛋白决定造血谱系中细胞命运。 PU.1 据报道，反式激活（TAD）和DNA结合结构域为参与互动，我们注意到PU.1 TAD 与p53的TAD共享结构同源性。与 Jim Omichinski的实验室我们已经表明，p53 TAD在体外和体内也与GATA-1 DNA结合结构域相互作用。互动需要GATA-1的接头和C指。蛋白质相互抑制在红细胞前体细胞系中彼此的反式激活活性，6C2。可能需要GATA-1来防止核凝结和摘除液的p53诱导，该核凝结和摘除率是在红细胞形成之前或巨核细胞多倍体化期间的。与Yamamoto Masi合作，我们正在测试GATA-1的突变体，这些突变体与p53在GATA1.05细胞和小鼠的救援测定中不相互作用，以确定这种相互作用在造血过程中的作用。另一个项目涉及与EKLF转录因子相关的贫血。 EKLF DNA结合结构域中的突变发生在贫血患者中。 CBA和NAN（新生儿贫血）小鼠中EKLF第二个锌指的氨基酸被突变。与吉姆·比克（Jim Bieker）合作，我们正在尝试确定某些EKLF靶基因的Nan-Eklf相对于野生型的Nan-Eklf的不同DNA结合亲和力是否基于不同的DNA结合亲和力。