Structure and Function of SET Domain Methyltransferases

SET结构域甲基转移酶的结构和功能

基本信息

批准号：
7571638
负责人：
RAYMOND C TRIEVEL
金额：
$ 25.22万
依托单位：
UNIVERSITY OF MICHIGAN AT ANN ARBOR
依托单位国家：
美国
项目类别：
财政年份：
2006
资助国家：
美国
起止时间：
2006-02-01 至 2011-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7571638
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

项目摘要

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.

组蛋白是组织和压缩染色质核 DMA 的主要支架蛋白。这些蛋白质经历多种类型的化学修饰，这些修饰控制着体内的各种过程核。氨基酸赖氨酸的甲基化发生在几种组蛋白中，并且与基因表达以及 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) 是催化组蛋白内特定赖氨酸甲基化的酶。 HKMT 已被证明可以作为基因表达的正调节因子和负调节因子，取决于它们在组蛋白中甲基化的特定赖氨酸。最近，一些HKMT被展示甲基化转录因子（激活基因表达的 DNA 结合蛋白），表明这些酶在控制转录方面的作用比以前认为的更广泛。的重要性这些酶与 HKMT 之间的直接关系强调了 HKMT 在调节基因表达方面的作用。多种形式的癌症。多个 HKMT 的缺失、突变或过度表达已被证实存在关联白血病、前列腺癌、乳腺癌、肺癌、结直肠癌和肝癌等恶性肿瘤。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导致癌症，我们提出了几个实验来表征它们的蛋白质底物特异性和阐明它们甲基化核蛋白中特定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，其中我们将确定这些酶与其蛋白质底物结合的分子结构。综合起来，这研究不仅将为细胞核中 HKMT 的功能提供新的见解，而且还将提供开发针对这些酶的特异性抑制剂来治疗癌症的途径。目的1：通过生化和结构确定HKMTs赖氨酸甲基化的化学机制对称为 Rubisco 大亚基甲基转移酶 (LSMT) 的植物 HKMT 同源物的研究。目标 2：确定人 SET7/9（一种组蛋白和转录因子）的底物特异性甲基转移酶，通过结构和生化技术。目标 3：确定人类 SETS 的结构和底物特异性，这是一种在调节中至关重要的 HKMT 细胞分裂过程中的基因沉默和适当的染色体分离。