Regulation of histone methylation

组蛋白甲基化的调控

• 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: 2014
• 资助国家: 加拿大
• 起止时间: 2014-01-01 至 2015-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=547990
• 关键词: Regulation; histone; methylation

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，染色质是一种由DNA、组蛋白和非组蛋白组成的核蛋白结构。染色质对基因表达的保真度很重要，其结构可以通过在组蛋白上添加翻译后修饰（PTMs）来调节。一种这样的修饰，赖氨酸36处组蛋白H3的甲基化，是通过RNA聚合酶II的传递来靶向活性基因的，这种PTM可以说是所有真核生物中最公认的高产转录标志。然而，尽管对H3K36甲基化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，人们对这种修饰如何在基因抑制后被去除知之甚少。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参与H3K36甲基化调控的基因保存良好，并且这种模式生物的遗传可追溯性。所使用的技术包括ChIP（染色质免疫沉淀）、qPCR（定量聚合酶链反应）、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资助的学生将学习各种各样的技能，这些技能在生命科学研究的多个领域受到高度追捧。