Structure and Function of SET Domain Methyltransferases

SET结构域甲基转移酶的结构和功能

• 批准号: 7933154
• 负责人: RAYMOND C TRIEVEL
• 金额: $ 13.38万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-30 至 2011-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7933154
• 关键词: Active Sites; Amino Acids; Binding; Binding Sites; Biochemical; Biochemistry; Biological Assay; Biological Process; Breast; Cell Cycle; Cell Cycle Checkpoint; Cell Division Process; Cell Nucleus; Cell division; Chemicals; Chromatin; Chromosomal Stability; Chromosome Segregation; Chromosomes; Colorectal; Complex; Cytokinesis; DNA Repair; DNA-Binding Proteins; Defect; Deletion Mutation; Development; Enzymes; Gene Expression; Gene Expression Regulation; Gene Silencing; Genetic Transcription; Goals; Heterochromatin; Histone H3; Histone H4; Histone-Lysine N-Methyltransferase; Histones; Homologous Gene; Human; Kinetics; Knowledge; Laboratories; Lead; Libraries; Link; Lung; Lysine; Malignant Neoplasms; Malignant neoplasm of liver; Measures; Methylation; Methyltransferase; Mitosis; Mitotic; Modeling; Modification; Molecular Structure; Mono-S; Mutation; Nuclear; Nuclear Protein; Nuclear Proteins; Peptides; Plants; Play; Process; Property; Prostate; Proteins; Reaction; Regulator Genes; Reporting; Research; Research Personnel; Ribulose-Bisphosphate Carboxylase; Role; SET Domain; Scaffolding Protein; Site; Specificity; Structure; Substrate Specificity; TAF10 gene; TP53 gene; Techniques; Transcriptional Activation; Transcriptional Regulation; Variant; amino group; analog; cancer therapy; enzyme structure; enzyme substrate; inhibitor/antagonist; insight; leukemia; novel; programs; research study; structural biology; transcription factor

Histones are the major scaffolding proteins which organize and compact nuclear DMAin chromatin. These proteins undergo numerous types of chemical modifications that govern a variety of processes in the nucleus. Methylation of the amino acid lysine occurs in several histones and has been linked to the regulation of gene expression as well as in DNA repair processes and cell division. Histone lysine methyltransferases (HKMTs) are enzymes that catalyze the methylation of specific lysines within histones. HKMTs have been shown to function as both positive and negative regulators of gene expression, depending on the specific lysine that they methylate in histones. Recently, some HKMTs have been shown to methylate transcription factors (DNA binding proteins that activate gene expression), suggesting that these enzymes have broader roles in controlling transcription than previously believed. The importance of HKMTs in regulating gene expression is underscored by direct relationships between these enzymes and numerous forms of cancer. Deletions, mutations, or over-expression of several HKMTs have been linked malignancies such as leukemias and prostate, breast, lung, colorectal, and hepatic cancers. In order to gain insight into the functions of HKMTs in transcriptional regulation and how disruption of these enzymes can lead to cancer, we propose several experiments to characterize their protein substrate specificities and elucidate the chemical mechanism through which they methylate specific lysine residues in nuclear proteins. To achieve these goals, we will use a combination of biochemistry and structural biology in which we will determine the molecular structures of these enzymes bound to their protein substrates. Taken together, this research will provide not only novel insights into the functions of HKMTs in the nucleus, but will also provide avenues for the development of specific inhibitors against these enzymes in the treatment of cancer. Aim1: Determine the chemical mechanism of lysine methylation of HKMTs via biochemical and structural studies of a plant HKMT homolog known as Rubisco large subunit methyltransferase (LSMT). Aim2: Determine the substrate specificity of human SET7/9, a histone and transcription factor methyltransferase, through structural and biochemical techniques. Aim3: Determine the structure and substrate specificity of human SETS, an HKMT that is pivotal in regulating gene silencing and proper chromosome separation during cell division.

组蛋白是组织和紧凑的核Dmain染色质的主要脚手架蛋白。这些 蛋白质经历了多种类型的化学修饰，这些化学修饰控制了各种过程 核。氨基酸赖氨酸的甲基化发生在几个组蛋白中，已与 基因表达以及DNA修复过程和细胞分裂的调节。组蛋白赖氨酸 甲基转移酶（HKMT）是催化组蛋白中特定赖氨酸的甲基化的酶。 HKMT已被证明是基因表达的阳性和阴性调节剂， 取决于它们在组蛋白中甲基化的特定赖氨酸。最近，一些HKMT已显示 甲基化转录因子（激活基因表达的DNA结合蛋白），表明 这些酶在控制转录中具有比以前认为的更广泛的作用。重要性 调节基因表达中的HKMT被这些酶与 多种形式的癌症。几个HKMT的删除，突变或过表达已链接 白血病和前列腺，乳房，肺，结直肠和肝癌等恶性肿瘤。为了获得 深入了解HKMT在转录调控中的功能以及这些酶的破坏如何可以 导致癌症，我们提出了几个实验，以表征其蛋白质底物特异性和 阐明了它们在核蛋白中甲基化特异性赖氨酸残基的化学机制。 为了实现这些目标，我们将结合生物化学和结构生物学的结合。 确定与其蛋白质底物结合的这些酶的分子结构。总的来说，这个 研究不仅将提供有关HKMT在核中功能的新见解，还将提供 针对癌症治疗的这些酶开发特异性抑制剂的途径。 AIM1：通过生化和结构确定HKMT的赖氨酸甲基化的化学机制 研究植物HKMT同源物，称为Rubisco大亚基甲基转移酶（LSMT）。 AIM2：确定人Set7/9（组蛋白和转录因子）的底物特异性 通过结构和生化技术，甲基转移酶。 AIM3：确定人类集合的结构和底物特异性，这是一个在调节的HKMT 细胞分裂期间的基因沉默和适当的染色体分离。