Studying the role of H3K36 methylation in development and gene expression

摘要 组蛋白的共价翻译后修饰(PTM)被广泛认为促进表观遗传 基因表达信息从一个细胞世代遗传到下一代细胞。然而，这一前提并没有 直接在动物身上进行测试，限制了我们对PTMS对复杂分子或 构成人类疾病基础的发育过程。为了审问这种关系，我们开发了一种 黑腹果蝇内源性组蛋白基因簇替换的遗传策略 五个核心组蛋白亚基的工程多基因阵列，在可修饰残基中具有特定的突变。 组蛋白H3亚基(H3K36)中赖氨酸36的甲基化是一种转录连锁的PTM，具有很好的 在抑制隐蔽起始和调节选择性剪接方面的特征作用，使其成为 使用我们的系统进行研究具有吸引力的选择。我们的目标是使用H3K36突变模型来评估这种贡献 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甲基化在全球基因表达调控中的分子要求和替代 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甲基化的分子机制之间的相互作用 与疾病相关的部分、细胞和发育过程。

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.