Role of Histone H3 Lysine 36 Methylation in Chromatin

组蛋白 H3 赖氨酸 36 甲基化在染色质中的作用

• 批准号: 7050183
• 负责人: Ronald Laribee
• 金额: $ 4.32万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-02-01 至 2006-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7050183
• 关键词: chromatin; chromatin immunoprecipitation; gene expression; gene induction /repression; genetic regulation; genetic regulatory element; histones; lysine; methylation; microarray technology; nucleic acid structure; postdoctoral investigator; protein binding; protein protein interaction; protein structure function; yeasts

DESCRIPTION (provided by applicant): Recent studies into the functions of histone methylation have revealed this histone modification to have many roles in gene regulation. In budding yeast, Set2 is the sole H3 lysine 36 methyltransferase. It functions to either mono-, di, or tri-methylate this residue. Methylation of lysine 36 has been associated both with gene repression and transcription elongation, yet how methylation functions in these roles is not understood. To determine the role of H3 lysine 36 methylation in regulating gene expression, a chromatin immunoprecipitation approach, coupled with microarray analysis, will be used to determine the genomic locations of the three different methyl forms of K36. A gene specific chromatin immunoprecipitation approach will be used subsequent to the microarray analysis to precisely define within the body of genes where the individual methylation marks are localized. Furthermore, proteins that bind to the different methylated forms of H3 K36 to mediate Set2's downstream effects on gene expression will be characterized and their role in regulating gene activity assessed. These experiments will further elucidate the functions of the different methyl K36 forms in regulating gene activity and chromatin function.

描述（由申请人提供）： 最近对组蛋白甲基化功能的研究表明，这种组蛋白修饰在基因调控中具有多种作用。在芽殖酵母中，Set2 是唯一的 H3 赖氨酸 36 甲基转移酶。它的作用是将该残基单甲基化、二甲基化或三甲基化。赖氨酸 36 的甲基化与基因抑制和转录延伸相关，但甲基化如何在这些作用中发挥作用尚不清楚。为了确定 H3 赖氨酸 36 甲基化在调节基因表达中的作用，染色质免疫沉淀方法与微阵列分析相结合，将用于确定 K36 三种不同甲基形式的基因组位置。微阵列分析后将使用基因特异性染色质免疫沉淀方法，以精确定义基因体内各个甲基化标记的定位位置。此外，将表征与 H3 K36 的不同甲基化形式结合以介导 Set2 对基因表达的下游影响的蛋白质，并评估它们在调节基因活性中的作用。这些实验将进一步阐明不同甲基K36形式在调节基因活性和染色质功能中的功能。