Studying the role of H3K36 methylation in development and gene expression

研究 H3K36 甲基化在发育和基因表达中的作用

基本信息

批准号：
8649283
负责人：
Michael P Meers
金额：
$ 2.97万
依托单位：
UNIV OF NORTH CAROLINA CHAPEL HILL
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-03-01 至 2017-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8649283
关键词：
Affect Alleles Alternative Splicing Animal Model Animals Apoptotic Caspase Cell Death Cell Proliferation Cell physiology Cells Charge Cleaved cell Complex DNA Data Defect Development Developmental Process Disease Drosophila melanogaster Embryo Engineering Enzymes Epigenetic Process Gene Cluster Gene Expression Gene Expression Regulation Generations Genetic Genetic Transcription Global Change Histone H3 Histones Immunofluorescence Immunologic Incidence Introns Laboratories Link Lysine Methylation Methyltransferase Mitosis Modeling Modification Molecular Morphology Mutation Nature Nonsense Codon Nonsense-Mediated Decay Organism Pathway interactions Pattern Phenotype Play Poly(A)+ RNA Population Post-Translational Protein Processing Process Proteins RNA Sequences RNA Splicing Regulation Renal Cell Carcinoma Role Severities Staging System Testing Time Tissues Transcript Transcription Initiation Yeasts disease phenotype gene replacement genome-wide histone modification human disease imaginal disc improved insight mRNA Precursor molecular phenotype mutant prevent programs public health relevance research study stem transcriptome sequencing

项目摘要

Abstract Covalent post-translational modifications (PTMs) of histone proteins are widely believed to facilitate epigenetic inheritance of gene expression information from one cell generation to the next. However, this premise has not been directly tested in animals, limiting our understanding of the contribution of PTMs to complex molecular or developmental processes that underlie human disease. To interrogate this relationship, we have developed a genetic strategy in Drosophila melanogaster for replacement of the endogenous histone gene cluster with an engineered multigene array of the five core histone subunits with specific mutations in modifiable residues. Methylation of lysine 36 in the histone H3 subunit (H3K36) is a transcription-linked PTM that has well- characterized roles in suppression of cryptic initiation and regulation of alternative splicing, making it an attractive choice for study using our system. We aim to use H3K36 mutant models to evaluate the contribution of H3K36 methylation to proper proliferation and development. Additionally, we aim to directly test the molecular requirement for H3K36 methylation in proper regulation of global gene expression and alternative splicing, and to correlate molecular role with observed cellular and developmental phenotypes. These studies will be the first to directly test the interplay between molecular mechanisms in which H3K36 methylation takes a part and cellular and developmental processes that are relevant to disease.

抽象的人们普遍认为组蛋白的共价翻译后修饰 (PTM) 有助于表观遗传基因表达信息从一代细胞遗传到下一代。然而这个前提并没有直接在动物身上进行测试，限制了我们对 PTM 对复杂分子或人类疾病背后的发育过程。为了探究这种关系，我们开发了一个果蝇内源组蛋白基因簇替换的遗传策略工程多基因阵列的五个核心组蛋白亚基具有可修饰残基的特定突变。组蛋白 H3 亚基 (H3K36) 中赖氨酸 36 的甲基化是一种转录相关 PTM，具有良好的在抑制隐秘起始和调节选择性剪接方面具有特征性的作用，使其成为使用我们的系统进行学习的有吸引力的选择。我们的目标是使用 H3K36 突变模型来评估贡献 H3K36 甲基化对正常增殖和发育的影响。此外，我们的目标是直接测试 H3K36 甲基化在正确调控全局基因表达和替代中的分子要求剪接，并将分子作用与观察到的细胞和发育表型相关联。这些研究将是第一个直接测试 H3K36 甲基化分子机制之间相互作用的人与疾病相关的部分、细胞和发育过程。