Detecting Histone Modification Turnover in Living Cells

检测活细胞中的组蛋白修饰更新

基本信息

批准号：
7927086
负责人：
Karmella Ann Haynes
金额：
$ 5.38万
依托单位：
HARVARD MEDICAL SCHOOL
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-08-01 至 2011-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7927086
关键词：
Acetylation Amino Acid Motifs Antineoplastic Agents Behavior Binding Biomedical Engineering Cell Cycle Cell Differentiation process Cell division Cells Chemicals Chromatin DNA Repair Development Disease Drosophila genus Drug Delivery Systems EZH2 gene Engineering Enzymes Epigenetic Process Feedback Fellowship Fluorescence Gene Activation Genes Genetic Genome Histone Code Histone H3 Histones Human In Vitro Individual Inherited Instruction Lead Length Life Lysine Mammalian Cell Maps Measures Mediating Memory Messenger RNA Methylation Mitosis Modification Mus Names Nuclear Output Pattern Peptides Phosphorylation Polycomb Program Development Protein Binding Proteins Reading Recombinants Reporter Reporting Repression Research Research Proposals Signal Transduction Silver Specificity Stimulus Synthetic Genes System Tail Technology Testing Time Transcriptional Activation Transgenic Organisms Translating Work Zebrafish base biological systems chromatin immunoprecipitation chromatin modification design fly gene repression histone methyltransferase histone modification in vivo mammalian genome protein structure research study synthetic biology

项目摘要

DESCRIPTION (provided by applicant): In the post-genome era, one of the current challenges is to understand how epigenetic mechanisms impact cellular programming, development and disease. Histones, the proteins around which DMA is wrapped, carry covalent modifications that lead to gene activation or repression. Effector proteins read and translate this "histone code" through histone binding motifs. Recent studies have generated a wealth of information on patterns of histone modifications. Current technologies to assess histone modifications rely upon chromatin immunoprecipitation (ChIP). While powerful, ChIP has limited ability to probe the dynamics of epigenetic modification in living cells. I propose to use a modular histone tail binding motif from the polycomb (Pc) group of effector proteins to measure changes in histone methylation at a single locus in living cells. Synthetic biology is a new experimental paradigm that offers creative opportunities to investigate the mechanisms of epigenetic inheritance. The design of artificial biological systems is a means for testing our understanding of natural protein "modules." To this end, I will identify a peptide from PC that acts as a module that specifically recognizes methylated histone tails in mammalian cells. I will integrate this new modular part into a reporter system designed to track changes in histone methylation in dividing cells. Peptides that read specific histone modifications could serve as a new class of modular parts that interface bioengineered gene circuits with epigenetic signals that mark key steps in cell development. Heritable changes in the proteins bound to DMA (epigenetics) underlie mechanisms of cell development and disease. Research of the enzymes that mediate epigenetic changes has lead to the discovery of new cancer drug targets. Here, we use a bioengineering approach to probe the epigenetic phenomena. PHS 416-1 (Rev.10/05) Page 2 Number pages consecutively at the bottom throughout Form Page 2 the application. Do not use suffixes such as 2a, 2b. NAME OFAPPLICANT (Last, first, middle initial) Kirschstein-NRSA Individual Fellowship Application Haynes, Karmella A. (To becompletedbyapplicant - follow PHS 416-1 instructions)

描述（由申请人提供）：在基因组后时代，目前的挑战之一是了解表观遗传机制如何影响细胞编程，发育和疾病。组蛋白，包裹DMA周围的蛋白质，带有共价修饰，从而导致基因激活或抑制。效应子蛋白通过组蛋白结合基序读取和翻译此“组蛋白代码”。最近的研究产生了有关组蛋白修饰模式的大量信息。当前评估组蛋白修饰的技术取决于染色质免疫沉淀（CHIP）。虽然功能强大，但CHIP具有有限的探测活细胞表观遗传修饰动力学的能力。我建议使用来自多肉液（PC）效应蛋白的模块化组蛋白尾巴结合基序来测量活细胞中单个基因座的组蛋白甲基化的变化。合成生物学是一种新的实验范式，提供了研究表观遗传遗传机制的创造机会。人工生物系统的设计是测试我们对天然蛋白“模块”的理解的一种手段。为此，我将从PC中确定一个肽，该肽充当一个模块，该模块专门识别哺乳动物细胞中的甲基化组蛋白尾巴。我将将这个新的模块化部分整合到一个旨在跟踪分隔单元中组蛋白甲基化的变化的报告系统中。读取特定组蛋白修饰的肽可以用作新的模块化部分，这些模块化部分将生物工程基因与表观遗传信号接触，这些信号标志着细胞发育的关键步骤。与DMA（表观遗传学）结合的蛋白质的遗传变化是细胞发育和疾病的基础。对介导表观遗传变化的酶的研究导致发现了新的癌症药物靶标。在这里，我们使用一种生物工程方法来探测表观遗传现象。 PHS 416-1（Rev.10/05）第2页数字页面在整个表2页上的底部连续底部连续2页。请勿使用后缀2a，2b。 Applicant的名称（最后，第一个，中间缩写）Kirschstein-nrsa个人奖学金申请Haynes，Karmella A.