CHROMATIN AND EPIGENETIC INHERITANCE

染色质和表观遗传

基本信息

批准号：
7429032
负责人：
Kathrin Plath
金额：
$ 231万
依托单位：
UNIVERSITY OF CALIFORNIA LOS ANGELES
依托单位国家：
美国
项目类别：
财政年份：
2007
资助国家：
美国
起止时间：
2007-09-30 至 2012-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7429032
关键词：
Accession Number Acetylation Adaptor Signaling Protein Address Affect Affinity Affinity Chromatography Agreement Alleles Amino Acids Ammonium Sulfate Anions Anterior Antibodies Antisense RNA Appearance Atomic Force Microscopy Automobile Driving Bacteriophages Binding Binding Sites Biochemical Biochemical Genetics Biochemistry Biological Biological Assay Biological Models Biology Biotin Books Boxing Breeding Caenorhabditis elegans Capsid Proteins Cations Cell Cycle Cell Cycle Regulation Cell Cycle Stage Cell Differentiation process Cell Extracts Cell Fraction Cell Line Cell Nucleus Cell division Cells Centrifugation Chemicals Chimeric Proteins Chromatin Chromatin Structure Chromatography Chromosomes Code Collaborations Complex Computer software Coupled Custom Cytolysis Cytomegalovirus Cytosine DNA DNA Methylation DNA Methyltransferase DNA Modification Methylases DNA Modification Process DNA Sequence DNA biosynthesis Data Data Set Detection Development Dissection Doctor of Philosophy Dosage Compensation (Genetics)Drosophila genus ES Cell Line Ectopic Expression Embryo Endopeptidase K Endoplasmic Reticulum Enterobacteria phage MS2 Enzymes Epigenetic Process Epitopes Eukaryota Eukaryotic Cell Excision Exons Failure Family member Female Fibroblasts Figs - dietary Fluorescence Microscopy Fluorescent Dyes Fluorescent in Situ Hybridization Fractionation Future Gene Expression Gene Expression Profile Gene Expression Regulation Gene Silencing Gene Targeting Generations Genes Genetic Genetic Recombination Genetic Transcription Genets Genome Genomic Imprinting Genomics Genotype Glass Green Fluorescent Proteins Hearing Heart Heterochromatin High Prevalence Histone Code Histone H3 Histones Homeobox Genes Hour Human Hybrids Image Imagery Immunofluorescence Microscopy Immunoprecipitation In Vitro Incubated Inherited Institutes Instruction Intention Knock-out Label Laboratories Lead Length Libraries Ligands Light Link Localized Location Lysine MS2 coat protein Maintenance Mammalian Cell Mammals Manuscripts Mass Spectrum Analysis Measurement Mediating Membrane Membrane Proteins Memory Methods Methylation Methyltransferase Microarray Analysis Micrococcal Nuclease Microscope Microscopic Minor Mitosis Mitotic Chromosome Modification Modification Type Molecular Analysis Monitor Mothers Mouse Strains Mus Mutation Nature Nocodazole Nuclear Extract Nuclear RNA Nucleosome Core Particle Nucleosomes Numbers Open Reading Frames Optics PRC1 Protein Pan Genus Pattern Peptide Signal Sequences Phase Phenotype Plant Resins Plasmids Play Polycomb Polymerase Chain Reaction Population Postdoctoral Fellow Preparation Procedures Process Protein Binding Protein Family Protein Overexpression Proteins Proteomics Publishing Puromycin RNA RNA Binding RNA Interference RNA Phages RNA Probes RNA Stability RNA purification RNA-Binding Proteins RNA-Protein Interaction Recombinants Recruitment Activity Regulation Reporter Reporter Genes Research Research Design Research Personnel Resistance Resolution Reverse Transcription Ribonucleoproteins Robot Role Sampling Scanning Scanning Probe Microscopes Scheme Science Screening procedure Sepharose Silicon Dioxide Site Small Interfering RNA Somatic Cell Source Southern Blotting Spectrometry Spermatogenesis Streptavidin Students Sucrose System Tail Technology Technology Transfer Testing Tetracycline Tetracyclines Thinking Thymidine Time Tobramycin Transcript Transfection Transferase Transgenes Translating Ubiquitination Undifferentiated Untranslated RNA Variant Work X Chromosome X Inactivation Yeasts absorption aptamer base chromatin immunoprecipitation chromatin protein chromatin remodeling crosslink daughter cell demethylation design embryonic stem cell experience fly gene repression genome-wide analysis histone methyltransferase homologous recombination human female hygromycin A in vitro Assay in vivo insight intracellular protein transport knock-down macroH2A histone male mammalian genome member mouse genome neglect novel novel strategies nuclear reprogramming null mutation numb protein programs promoter protein crosslink protein localization location recombinase reconstitution repaired research study secretory protein single molecule size small hairpin RNA stem synergism tandem mass spectrometry technology development tool transcription factor transmission process

项目摘要

Covalent modifications of both DNA and histones are important for regulating gene expression. Here, I propose to address two fundamental, unanswered questions about chromatin modifications: i) how are changes in chromatin states established during development; and ii) once established, how are chromatin modifications stably preserved through future cell divisions? Initially, we will focus our analysis on understanding how the noncoding RNA Xist establishes silencing of the X chromosome in female mammalian cells. During initiation of X- inactivation, Xist RNA spreads in cis to coat the X chromosome that will become inactive, mediates silencing, and triggers the sequential accumulation of chromatin modifications. How Xist RNA initiates silencing remains unknown. One approach to gain insight into the function of Xist is to identify interacting proteins. We have obtained results demonstrating that Xist is part of a large protein complex. In Aim 1, we therefore propose to use classical and non- conventional purification strategies to identify Xist -interacting proteins. To identify proteins necessary for maintaining the silence of the inactive X, we will perform an RNAi based screen in Aim 2, which is based on the reactivation of the inactive X. We expect to find proteins involved in the epigenetic inheritance of the silent X chromosome state. The cell cycle poses a particularly challenging problem for epigenetic inheritance since histone modifications have to be maintained when DNA strands are duplicated during S phase. A major question therefore is, how chromatin modifications are transmitted through cell divisions, and if they are indeed sufficient as carriers of the epigenetic information. Surprisingly, we have found that histone modifications on the inactive X do not accumulate throughout the cell cycle. In Aim 3, we will extent studies on the cell cycle regulation of histone modifications.

DNA 和组蛋白的共价修饰对于调节基因表达非常重要。在这里，我建议解决有关染色质的两个基本的、尚未解答的问题修改：i) 发育过程中染色质状态的变化是如何发生的；和 ii) 一旦建立，染色质修饰如何在未来的细胞中稳定保存部门？首先，我们的分析重点是了解非编码 RNA 是如何存在的建立雌性哺乳动物细胞中 X 染色体的沉默。在X-启动期间失活，Xist RNA 以顺式扩散，覆盖 X 染色体，从而变得失活，介导沉默，并触发染色质修饰的连续积累。如何 Xist RNA 启动沉默仍然未知。深入了解功能的一种方法 Xist 是为了识别相互作用的蛋白质。我们获得的结果证明 Xist 是一个大的蛋白质复合物。因此，在目标 1 中，我们建议使用经典和非鉴定 Xist 相互作用蛋白的常规纯化策略。鉴定蛋白质为了维持非活性 X 的沉默，我们将进行基于 RNAi 的筛选目标 2，基于失活 X 的重新激活。我们期望找到相关蛋白质沉默X染色体状态的表观遗传。细胞周期构成了表观遗传的特别具有挑战性的问题，因为组蛋白修饰必须当 DNA 链在 S 期复制时得以维持。因此，一个主要问题是，染色质修饰如何通过细胞分裂传递，以及它们是否确实如此足以作为表观遗传信息的载体。令人惊讶的是，我们发现组蛋白对非活性 X 的修饰不会在整个细胞周期中累积。在目标 3 中，我们将组蛋白修饰对细胞周期调控的深入研究。

项目成果

期刊论文数量（13）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

A high-throughput screen of inactive X chromosome reactivation identifies the enhancement of DNA demethylation by 5-aza-2'-dC upon inhibition of ribonucleotide reductase.

DOI：
10.1186/s13072-015-0034-4
发表时间：
2015
期刊：
Epigenetics & chromatin
影响因子：
3.9
作者：
Minkovsky A;Sahakyan A;Bonora G;Damoiseaux R;Dimitrova E;Rubbi L;Pellegrini M;Radu CG;Plath K
通讯作者：
Plath K

Long-range chromatin contacts in embryonic stem cells reveal a role for pluripotency factors and polycomb proteins in genome organization.

DOI：
10.1016/j.stem.2013.08.013
发表时间：
2013-11-07
期刊：
CELL STEM CELL
影响因子：
23.9
作者：
Denholtz, Matthew;Bonora, Giancarlo;Chronis, Constantinos;Splinter, Erik;de Laat, Wouter;Ernst, Jason;Pellegrini, Matteo;Plath, Kathrin
通讯作者：
Plath, Kathrin

X chromosome reactivation in reprogramming and in development.

X 染色体在重编程和发育中重新激活。