Roles of the methylase NSD3 in neural crest migration and non-histone methylation

甲基化酶 NSD3 在神经嵴迁移和非组蛋白甲基化中的作用

• 批准号: 9259108
• 负责人: Jeffrey Barcus
• 金额: $ 2.98万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-09-02 至 2019-09-01
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9259108
• 关键词: Adrenal Medulla; Amino Acids; Animals; Biological Assay; Brain; Cartilage; Cell Culture System; Cell Culture Techniques; Cells; Chick Embryo; Congenital Abnormality; Cytoplasmic Protein; Data; Defect; Development; Disease; Embryo; Event; Face; Gene Expression; Genes; Goals; Heart; Histone H3; Histones; Human; In Situ Hybridization; Kinetics; Label; Lead; Link; Lysine; Malignant Neoplasms; Mass Spectrum Analysis; Mediating; Melanoma Cell; Messenger RNA; Metastatic Melanoma; Methylation; Methyltransferase; Modeling; Molecular; Multipotent Stem Cells; Neoplasm Metastasis; Neural Crest; Neural Crest Cell; Neuroblastoma; Neurons; Outcome; Pathway interactions; Peripheral Nervous System; Phenotype; Play; Population; Post-Translational Protein Processing; Prevention; Protein Methylation; Proteins; Proteomics; Regulation; Research; Role; Spinal Cord; Stable Isotope Labeling; Stem cells; Structure; Time; Transplantation; Vertebral column; Western Blotting; Work; base; bone; cancer therapy; cell behavior; cell motility; cell type; cleft lip and palate; craniofacial; craniofacial development; histone methyltransferase; in vivo; innovation; insight; knock-down; live cell imaging; loss of function; melanocyte; melanoma; migration; mutant; non-histone protein; novel; orofacial cleft; prevent; research study; targeted treatment; vertebrate embryos

PROJECT SUMMERY/ABSTRACT: Neural crest cells, a multipotent stem cell-like population unique to vertebrate embryos, migrate great distances during development to form a wide variety of different cell types including craniofacial cartilage and bone, melanocytes, the adrenal medulla, and much of the peripheral nervous system. Defects in neural crest cell migration result in craniofacial maladies such as orofacial clefts, and dysregulation of neural crest derived cell types leads to a number of invasive cancers such as neuroblastoma. Recent work in the Gammill lab identified NSD3 as the first lysine methyltransferase that is essential for neural crest gene expression and migration. Although the known role of NSD3 is to dimethylate the 36th lysine residue of histone H3, depletion of NSD3 does not significantly alter H3K36me2 occupancy on neural crest genes even though these genes are no longer expressed. These results, as well as the cytoplasmic localization of NSD3 in migrating neural crest cells, the importance of cytoplasmic protein methylation during neural crest migration, and increasing evidence that “histone” methyltransferases can methylate additional non-histone proteins, leads to the hypothesis that NSD3 methylates non-histone targets in order to regulate neural crest cell migration and gene expression. To explore this hypothesis, this proposal aims to define the essential role of NSD3 during cell migration in cell culture and in chick embryos by characterizing NSD3 non-histone targets and the migratory features regulated by their methylation. This will be accomplished by (1) defining the specific aspects of migration that are defective in NSD3 knockdown cells, (2) identifying non-histone proteins methylated by NSD3 through mass spectrometry-based quantitative proteomics, and (3) assaying the requirement for NSD3 non-histone targets during migration, and determining the importance of NSD3-dependent methylation for their function. Accomplishing these goals will define a novel mode of regulation governing neural crest cell migration and craniofacial development, and will reveal new functions and targets of NSD3. These results will reveal unappreciated post-translational control over cell migration, will uncover a novel regulatory mechanism in development, and will have important implications for efforts to prevent craniofacial birth defects and develop treatments for metastasis.

夏季/摘要： 神经rest细胞是脊椎动物胚胎独有的多能干细胞样种群，迁移很大 发育过程中的距离形成各种不同的细胞类型，包括颅面软骨和 骨，黑色素细胞，肾上腺髓质和大部分周围神经系统。神经rest中的缺陷 细胞迁移导致颅面疾病（例如口腔裂缝）和神经rest的失调 细胞类型导致许多侵入性癌症，例如神经母细胞瘤。 Gammil Lab的最新工作 鉴定为NSD3是第一个对于神经rest基因表达和 迁移。尽管NSD3的已知作用是将组蛋白H3的第36-赖氨酸居住 NSD3并未显着改变神经rest基因上H3K36Me2的占有率，即使这些基因是 不再表达。这些结果以及NSD3在迁移神经crest中的细胞质定位 细胞，神经rest迁移过程中细胞质蛋白甲基化的重要性，越来越多的证据 “组蛋白”甲基转移酶可以甲基化附加的非固定蛋白，从而导致假设是 NSD3甲基化靶标的甲基化靶标，以调节神经rest细胞迁移和基因表达。到 探讨这一假设，该提案旨在定义NSD3在细胞迁移中的重要作用 通过表征NSD3非历史目标和迁移特征，培养和小鸡胚胎中的胚胎 通过它们的甲基化。这将通过（1）定义迁移的特定方面来实现 NSD3敲低细胞中有缺陷，（2）鉴定由NSD3通过质量甲基化的非固定蛋白 基于光谱法的定量蛋白质组学，以及（3）分析NSD3非固件目标的要求 在迁移过程中，并确定NSD3依赖性甲基化对其功能的重要性。 实现这些目标将定义一种新型的管理神经rest细胞迁移和 颅面发育，并将揭示NSD3的新功能和目标。这些结果将揭示 未经批准的翻译后控制细胞迁移，将发现一种新的调节机制 发展，将对防止颅面出生缺陷和发展的努力具有重要意义 转移的治疗方法。