Regulation of histone methylation

组蛋白甲基化的调控

基本信息

批准号：
262102-2013
负责人：
Howe, LeAnn
金额：
$ 2.62万
依托单位：
University of British Columbia
依托单位国家：
加拿大
项目类别：
Discovery Grants Program - Individual
财政年份：
2015
资助国家：
加拿大
起止时间：
2015-01-01 至 2016-12-31
项目状态：
已结题

来源：
https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=571238
关键词：
Regulation histone methylation

项目摘要

In eukaryotes DNA is packaged into chromatin, a nucleoprotein structure consisting of DNA, histones, and non-histone proteins. Chromatin is important for the fidelity of gene expression and its structure can be regulated by the addition of post-translational modifications (PTMs) to histones. One such modification, methylation of histone H3 at lysine 36, is targeted to active genes by the passage of RNA polymerase II, and this PTM is arguably the most recognized hallmark of productive transcription in all eukaryotes. However, despite over a decade worth of research into the mechanisms behind the establishment and function of H3K36 methylation, there is surprisingly little known about how this modification is removed following gene repression. The research proposed in this application will reveal the enzymes and pathways involved in removal of H3K36 methylation during gene repression. We will use S. cerevisiae as a model organism due to the excellent conservation of genes involved in regulation of H3K36 methylation between yeast and other eukaryotes, and the genetic tractability of this model organism. The techniques used include ChIP (chromatin immunoprecipitation), qPCR (quantitative polymerase chain reaction), library preparation for deep sequencing, bioinformatics, protein purification, enzyme kinetics, and yeast genetics, and thus students supported by my NSERC grant will learn a diverse set of skills that are highly sought after in multiple fields of life science research.

在真核生物中，DNA 被包装成染色质，这是一种由 DNA、组蛋白和非组蛋白组成的核蛋白结构。染色质对于基因表达的保真度非常重要，其结构可以通过向组蛋白添加翻译后修饰 (PTM) 来调节。其中一种修饰是组蛋白 H3 在赖氨酸 36 处的甲基化，通过 RNA 聚合酶 II 的通道靶向活性基因，并且这种 PTM 可以说是所有真核生物中最受认可的高效转录标志。然而，尽管对 H3K36 甲基化的建立和功能背后的机制进行了十多年的研究，但令人惊讶的是，人们对基因抑制后如何消除这种修饰知之甚少。本申请中提出的研究将揭示基因抑制过程中参与去除 H3K36 甲基化的酶和途径。我们将使用酿酒酵母作为模式生物，因为酵母和其他真核生物之间涉及 H3K36 甲基化调节的基因具有出色的保守性，并且该模式生物的遗传易处理性。使用的技术包括 ChIP（染色质免疫沉淀）、qPCR（定量聚合酶链式反应）、深度测序文库制备、生物信息学、蛋白质纯化、酶动力学和酵母遗传学，因此，由我的 NSERC 资助支持的学生将学习各种不同的技术在生命科学研究的多个领域中备受追捧的技能。