Function of a Putative Determinant in Hematopoiesis

造血作用中推定决定因素的功能

基本信息

批准号：
8074442
负责人：
JAMES J BIEKER
金额：
$ 34.83万
依托单位：
ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
依托单位国家：
美国
项目类别：
财政年份：
1993
资助国家：
美国
起止时间：
1993-08-01 至 2012-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8074442
关键词：
Acetylation Address Adult Antibodies Architecture Binding Biochemical Biological Assay Biological Process Biology Cell Line Cell Nucleus Cells Chromatin Chromatin Remodeling Factor Chromatin Structure Complex Cytoplasm DNA Data E1A-associated p300 protein Elements Environment Epigenetic Process Erythroid Erythroid Cells Erythropoiesis Fetal Liver Gene Activation Gene Expression Gene Expression Regulation Generations Genetic Genetic Transcription Globin Health Hematopoiesis Hematopoietic System Hepatocyte Histone H3 Investigation Lead Location Mediating Megakaryocytes Modification Molecular Mutation Nature Nuclear Nucleosomes Pattern Play Post-Translational Protein Processing Process Property Proteins Protocols documentation Regulation Regulator Genes Repression Repressor Proteins Role SMARCA4 gene Site Structure Suggestion System TAF9 gene Time Tissues Transactivation Transcription Initiation Transcriptional Activation gene repression in vivo interest novel progenitor programs promoter protein complex protein protein interaction research study transcription factor

项目摘要

DESCRIPTION (provided by applicant): Cell-restricted transcriptional modulators play critical roles in the process of selective gene regulation during hematopoiesis. We have been investigating the molecular and biological function of Erythroid Kr|ppel-like Factor (EKLF). EKLF is a cell-restricted transcription factor that is essential for completion of the erythroid program. Transcriptional activation is accomplished by its interaction with and modification by coactivators that lead to changes in chromatin structure and the onset of transcription, a process best established at the adult ?-globin locus. We find that EKLF also interacts with components of the nucleosome and of the transcriptional initiation complex, and propose in Aim 1 to illuminate how these varied interactions are integrated and properly assembled, leading to epigenetic modifications as a result of EKLF's actions. However, EKLF's general role is becoming increasingly more complex, as it is also expressed in the megakaryocyte-erythroid progenitor and interacts with two types of corepressors to repress transcription under constrained cell environments. Acetylation and sumoylation of EKLF play critical roles in the protein-protein contacts that are important for these effects, and the experiments of Aim 2 will decipher how these interactions are manifested within native repression targets. Finally, significant levels of EKLF reside in the cytoplasm of the erythroid cell in a form that shows subtle biochemical and functional differences compared to its nuclear version. The experiments of Aim 3 will address whether EKLF plays a role outside of the nucleus by isolating and characterizing cytoplasmic EKLF complexes. These studies will be aided by the use of in vivo chromatin binding assays and EKLF rescue systems in primary or minimally manipulated cells. Elucidating EKLF's role in regulatory phenomena will continue to illuminate novel aspects of erythroid biology and the essential mechanisms by which a cell-restricted transcription factor can exert varied yet highly controlled influences on expression that lead to genetic and epigenetic changes. PUBLIC HEALTH RELEVANCE: This proposal focuses on a continuing investigation of EKLF structure/function and how its post- translational modifications, protein-protein interactions, and protein-DNA interactions relate to its ability to generate erythroid cell-specific control of gene expression.

描述（由申请人提供）：细胞限制的转录调节剂在造血期间选择性基因调节过程中起关键作用。我们一直在研究红系KR | PPEL样因子（EKLF）的分子和生物学功能。 EKLF是一种细胞限制的转录因子，对于完成红细胞程序至关重要。转录激活是通过与共激活因子的相互作用和修饰来实现的，从而导致染色质结构变化和转录发作，这是在成人？ - 格珠蛋白基因座最佳建立的过程。我们发现EKLF还与核小体和转录起始复合物的组成部分相互作用，并提出AIM 1的建议，以阐明如何整合这些不同的相互作用并正确地组装这些相互作用，从而导致EKLF的作用导致表观遗传修饰。但是，EKLF的一般作用变得越来越复杂，因为它也在巨核细胞 - 雌雄同体祖细胞中表达，并与两种类型的Corepressor相互作用，以在约束细胞环境下抑制转录。 EKLF的乙酰化和Sumoylation在蛋白质 - 蛋白质接触中起关键作用，对这些影响很重要，AIM 2的实验将解读这些相互作用如何在天然抑制靶标内表现出来。最后，与其核版本相比，具有微妙的生化和功能差异的形式存在着显着水平的EKLF在红细胞细胞的细胞质中。 AIM 3的实验将通过隔离和表征细胞质EKLF复合物来解决EKLF是否在核之外起作用。这些研究将通过在原代或微小操纵细胞中使用体内染色质结合测定法和EKLF救援系统来帮助这些研究。阐明EKLF在调节现象中的作用将继续阐明红斑生物学的新颖方面，以及细胞限制的转录因子可以发挥不同但高度控制对表达的影响的基本机制，从而导致遗传和表观遗传变化。公共卫生相关性：该提案着重于对EKLF结构/功能的持续研究，以及其转化后修饰，蛋白质 - 蛋白质相互作用和蛋白-DNA相互作用如何与其产生基因表达的ery骨细胞特异性控制的能力有关。