Identifying non-histone targets of the neural crest methyltransferase NSD3

识别神经嵴甲基转移酶 NSD3 的非组蛋白靶点

• 批准号: 8645888
• 负责人: Daniel A. Kretzschmar
• 金额: $ 2.9万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-08-27 至 2016-08-26
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8645888
• 关键词: Amino Acids; Area; Binding; Biochemical; Biochemistry; Biological Assay; Cartilage; Catalytic Domain; Cell Culture Techniques; Cells; Complex; Congenital Abnormality; Craniofacial Abnormalities; Data; Defect; Development; Developmental Biology; Disease; Disseminated Malignant Neoplasm; Electroporation; Embryo; Endocrine; Ensure; Evaluation; Folate; Foundations; Gene Expression; Genetic; Goals; Health; Heart; Histone H3; Histones; Human; Immigration; In Situ Hybridization; In Vitro; Incidence; Investigation; Label; Length; Link; Lysine; Malignant Neoplasms; Mediating; Melanoma Cell; Mentors; Methylation; Methyltransferase; Molecular Target; Movement; Mutate; Nervous system structure; Neural Crest; Neural Crest Cell; Neuroblastoma; Neurons; Nuclear; Pathway interactions; Peripheral; Population; Post-Translational Protein Processing; Prevention; Process; Protein Array; Proteins; Proteomics; Regulation; Research; Role; SET Domain; Stable Isotope Labeling; Supplementation; Tail; Testing; Training; Travel; Waardenburg syndrome; Work; base; bone; cancer cell; cancer prevention; cancer therapy; cancer type; career; cell motility; cell type; cleft lip and palate; cofactor; craniofacial; design; experience; face skin; histone methyltransferase; improved; in vivo; insight; knowledge base; loss of function; melanocyte; melanoma; migration; multipotent cell; mutant; novel; prenatal; programs; public health relevance; screening; spatiotemporal

DESCRIPTION (provided by applicant): Neural crest cells are a population of vertebrate multipotent cells that migrate over long distances, form an impressive array of cell types, and are defective in diverse craniofacial abnormalities. Prenatal supplementation with folate, a cofactor in the methylation cycle, can reduce the incidence of craniofacial birth defects, yet unti recently any mechanistic impact of methylation on neural crest development was unknown. Recent data in the Gammill lab shows that the lysine methyltransferase NSD3 is required for neural crest cell migration. Based upon the cytoplasmic localization of both NSD3 and lysine methylated proteins in migratory neural crest cells, as well as increasing appreciation that 'histone' methyltransfserases methylate diverse non- histone proteins, this proposal postulates that NSD3-catalyzed methylation of non-histone proteins regulates neural crest cell migration. To investigate this possibility, the objective is to determine the importance of NSD3- mediated non-histone protein methylation during chick neural crest migration by identifying non-histone targets of NSD3 through two independent, complementary approaches in both directed and open-ended frameworks. This objective will be attained by: (1) screening NSD3-catalyzed methylation of candidate targets and arrayed proteins using an in vitro methyltranferase assay established by the candidate, (2) profiling NSD3-dependent non-histone protein methylation in C8161 melanoma cells using proteomics; and (3) determining the requirement for NSD3 target methylation during neural crest migration in vivo using chick transient genetic approaches. Achieving these aims will create a direct functional link between NSD3 non-histone methylation and neural crest cell migration, and will open exciting new avenues to explore in the treatment of aggressive cancers and the prevention of birth defects. Moreover, in completing this project, the candidate will acquire diverse expertise in proteomics, biochemistry, and developmental biology, giving him a broad knowledge base and a strong foundation in research. Mentoring by both a Sponsor and Co-Sponsor with extensive experience in these areas will ensure a successful completion of the research plan, as well as the implementation of a personalized training plan designed to prepare the candidate for a successful scientific career.

描述(申请人提供)：神经脊细胞是一群脊椎动物的多能细胞，可以远距离迁移，形成一系列令人印象深刻的细胞类型，并在各种颅面畸形中存在缺陷。产前补充叶酸是甲基化循环中的一个辅助因素，可以减少头面部出生缺陷的发生率，但最近甲基化对神经脊发育的任何机制影响尚不清楚。Gamill实验室的最新数据显示，赖氨酸甲基转移酶NSD3是神经脊细胞迁移所必需的。基于NSD3和赖氨酸甲基化蛋白在迁移的神经脊细胞中的细胞质定位，以及越来越多的人认识到组蛋白甲基转移酶甲基化不同的非组蛋白，这一假设假设NSD3催化的非组蛋白甲基化调节神经脊细胞的迁移。为了研究这种可能性，目的是通过在定向和开放式框架中通过两种独立的、互补的方法识别NSD3的非组蛋白靶标，来确定NSD3介导的非组蛋白甲基化在鸡神经脊迁移过程中的重要性。这一目标将通过以下方式实现：(1)使用候选人建立的体外甲基转移酶试验筛选NSD3催化的候选靶标和排列蛋白的甲基化；(2)使用蛋白质组学分析C8161黑色素瘤细胞中依赖NSD3的非组蛋白甲基化；以及(3)使用鸡瞬时遗传方法确定体内神经脊迁移过程中对NSD3靶标甲基化的需求。实现这些目标将在NSD3非组蛋白甲基化和神经脊细胞迁移之间建立直接的功能联系，并将为探索侵袭性癌症的治疗和预防出生缺陷开辟令人兴奋的新途径。此外，通过完成这个项目，候选人将获得蛋白质组学、生物化学和发育生物学方面的不同专业知识，使他拥有广泛的知识基础和坚实的研究基础。在这些领域拥有丰富经验的赞助者和共同赞助者的指导将确保成功完成研究计划，并实施旨在为候选人成功的科学生涯做准备的个性化培训计划。