Role of Histone H3 Lysine 36 Methylation in Chromatin

描述（由申请人提供）： 近年来对组蛋白甲基化功能的研究表明，组蛋白甲基化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。其功能是使该残基单甲基化、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长有关，但甲基化在这些作用中的作用尚不清楚。为了确定H3赖氨酸36甲基化在调节基因表达中的作用，将使用染色质免疫沉淀方法结合微阵列分析来确定K36的三种不同甲基形式的基因组位置。基因特异性染色质免疫沉淀方法将在微阵列分析之后使用，以精确地确定个体甲基化标记定位在基因体内的位置。此外，蛋白质结合到不同的甲基化形式的H3 K36介导Set2的下游效应的基因表达的特点和他们的作用，在调节基因活性的评估。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。

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.