Function of 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：
10681324
负责人：
JAMES J BIEKER
金额：
$ 64.29万
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 2025-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10681324
关键词：
ATAC-seq Acetylation Acylation Address Adult Affect Anemia Arginine Biochemical Biological Process Biology Blood Cells Butyrates Carnitine Cell Line Cells Chromatin Chromatin Structure Collaborations Consultations DNA DNA metabolism Data Enzymes Epigenetic Process Erythroid Erythroid Cells Erythropoiesis Event Fetal Hemoglobin Funding Gene Activation Gene Expression Regulation Genes Genetic Genetic Models Genetic Transcription Hand Hematopoiesis Heterozygote Histone Deacetylase Inhibitor Histones Human Human Cell Line Investigation Letters Link Longitudinal Studies Maps Metabolic Metabolism Methodology Modeling Modification Molecular Mus Mutation Nature Patients Phase Phosphorylation Play Post-Translational Protein Processing Process Property Protocols documentation Reagent Regulator Genes Role Series Site Sumoylation Pathway Testing Transcription Initiation Ubiquitination ascorbate beta Globin beta-Hydroxybutyrate erythroid Kruppel-like factor experimental study follow-up genetic approach histone modification in vivo induced pluripotent stem cell innovation interest metabolome metabolomics mutant novel programs promoter protein protein interaction success transcription factor

项目摘要

SUMMARY 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 Krüppel-like Factor (EKLF; KLF1). EKLF is a cell-restricted transcription factor that is a global regulator of genes essential for the erythroid program. Our proposal builds on unanticipated novel observations made during the previous funding cycle that extend this long-term study towards innovative directions: We found that EKLF is modified at arginine residues by mono- and di-methylation. The experiments of Aim 1 will address the context for these alterations and their function, and identify the enzymes responsible. We found that the metabolomes of the mouse Nan-EKLF and human CDA-KLF1 mutant erythroid cells are altered; this includes a metabolite involved in chromatin-associated changes in histone modification. The experiments of Aim 2 will examine the global nature of these modifications in mouse primary cells and in our newly-developed CDA patient-derived cell lines. We find that chromatin accessibility is altered in EKLF KO and in the Nan/+ cell. The experiments of Aim 3 will examine these changes in the context of the sets of decreased and ectopically expressed genes in these genetic backgrounds, an analysis that will be expanded to include CDA patient-derived cells. We found that the metabolome of the EKLF KO cell is altered, and leads to changes in DNA epigenetics. The experiments of Aim 4 will globally analyze these changes in the context of the altered RNA expression and chromatin accessibility observed in the absence of EKLF. These studies will be aided by the use of molecular, metabolic, and biochemical analyses, genetic approaches, and use of primary or minimally manipulated cells that have been newly established. 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 genetic expression, metabolism, and epigenetics.

概括细胞限制的转录调节剂在选择性基因的过程中起关键作用造血期间的调节。我们一直在研究红斑krüppel样因子（EKLF; KLF1）。 EKLF是一种细胞限制的转录因子，是全局红斑程序必不可少的基因调节子。我们的提议建立在意外的小说的基础上在先前的资金周期中进行的观察结果将这项长期研究扩展到创新方向：我们发现EKLF通过单甲基化和二甲基化在精氨酸残留物上进行了修饰。这 AIM 1的实验将解决这些更改及其功能的上下文，并确定负责的酶。我们发现小鼠NAN-EKLF和人CDA-KLF1突变体的代谢组细胞改变；这包括参与组蛋白染色质相关变化的代谢产物修改。 AIM 2的实验将检查这些修改的全球性质小鼠原代细胞以及我们新开发的CDA患者衍生的细胞系。我们发现在EKLF KO和NAN/+细胞中，染色质可及性发生了变化。这 AIM 3的实验将在下降的背景下检查这些变化，在这些遗传背景中以生态表达的基因，该分析将扩展到包括CDA患者衍生的细胞。我们发现EKLF KO细胞的代谢组发生了变化，并导致DNA变化表观遗传学。 AIM 4的实验将在全球范围内分析这些变化在没有EKLF的情况下观察到的RNA表达和染色质可及性改变了。这些研究将通过使用分子，代谢和生化分析的帮助来帮助遗传方法以及新的或最小操纵的细胞的使用阐明EKLF在监管现象中的作用将继续阐明红细胞生物学和细胞限制转录因子可以执行的基本机制对遗传表达，代谢和表观遗传学的影响多样化但高度控制的影响。

项目成果

期刊论文数量（51）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Blood group antigens reveal their maker.

血型抗原揭示了它们的制造者。

DOI：
10.1182/blood-2008-06-160580
发表时间：
2008
期刊：
Blood
影响因子：
20.3
作者：
Bieker,JamesJ
通讯作者：
Bieker,JamesJ

Functional interactions between erythroid Krüppel-like factor (EKLF/KLF1) and protein phosphatase PPM1B/PP2Cβ.

红系 Krüppel 样因子 (EKLF/KLF1) 和蛋白磷酸酶 PPM1B/PP2Cβ 之间的功能相互作用。

DOI：
10.1074/jbc.m112.350496
发表时间：
2012
期刊：
The Journal of biological chemistry
影响因子：
0
作者：
Yien,YvetteY;Bieker,JamesJ
通讯作者：
Bieker,JamesJ

Alternative splicing of EKLF/KLF1 in murine primary erythroid tissues.

小鼠原代红系组织中 EKLF/KLF1 的选择性剪接。

DOI：
10.1016/j.exphem.2014.08.007
发表时间：
2015
期刊：
Experimental hematology
影响因子：
2.6
作者：
Yien,YvetteY;Gnanapragasam,MerlinNithya;Gupta,Ritama;Rivella,Stefano;Bieker,JamesJ
通讯作者：
Bieker,JamesJ

GATA2 finds its macrophage niche.

GATA2 找到了它的巨噬细胞利基。