SAM Domains

SAM 域

• 批准号: 8053724
• 负责人: JAMES U BOWIE
• 金额: $ 22.32万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-04-01 至 2013-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8053724
• 关键词: Ankyrins; Autistic Disorder; Binding; Biological; Biological Assay; Biology; Blindness; Budgets; Cell physiology; Chromatography; Cleft Palate; Communication; Complex; DHFR gene; DNA Binding Domain; Diacylglycerol Kinase; Disease; Disease Pathway; Electron Microscopy; Electrons; Eukaryotic Cell; Family; Fibronectins; Funding; Gel; Gel Chromatography; Glutamine; Grant; Hereditary Disease; Homo; Human; Human Genome; Immune System Diseases; Investigation; Knowledge; Left; Leucine Zippers; Malignant neoplasm of brain; Medical; PH Domain; Phosphotransferases; Play; Polymers; Positioning Attribute; Protein Binding; Proteins; RNA; Role; SAM Domain; Scaffolding Protein; Solubility; Structure; System; Time; Transmembrane Domain; United States National Institutes of Health; Work; Zinc; biological systems; copolymer; deafness; experience; follow-up; genetic regulatory protein; leukemia; macromolecule; protein protein interaction; public health relevance; transcription factor

DESCRIPTION (provided by applicant): Sterile Alpha Motif (SAM) domains are among the most common protein modules in eukaryotic cells. They are found in many key regulatory proteins, scaffolding proteins, and transcription factors, which include many proteins involved in genetic diseases. Focused investigation of isolated systems has revealed that SAM domains are versatile protein-protein interaction partners, forming homo-oligomers, hetero-oligomers, homo- polymers, hetero-polymers and complexes with other macromolecules. Nevertheless, the vast majority of SAM domains remain functionally uncharacterized, leaving major gaps in our understanding of important biological pathways and disease states. Here we seek to leverage the considerable expertise we have developed with SAM domain structure and function to globally illuminate the functions of SAM domains in the human genome. The aims are: Aim I: Identify human SAM polymers. Every human-SAM domain will be expressed as a fusion to supercharged GFP to maintain polymer solubility. The SAM domains will be screened for aggregate formation by native gels and/or gel filtration chromatography. If they are found to form aggregates, they will be further examined by electron microscopy or AFM to distinguish aggregates from true polymers. Aim II: Identify all human homo- and hetero-SAM interactions discover polymer regulators and new copolymers. We will use a split DHFR assay to identify SAMs that bind to one another. Identified interactions will be verified biochemically with our GFP fusions from Aim I. We also hope to discover new SAM polymer regulators and possibly SAM copolymers. We believe the discovery-oriented approach proposed here is an efficient way to maximize the utility of our prior NIH funded advances for the benefit of others. PUBLIC HEALTH RELEVANCE: Cellular functions require precise communication between proteins. Disease occurs when these exchanges go awry. This project seeks to identify and characterize the interactions of a protein binding module found in about a hundred human proteins, including proteins directly involved in autism, leukemia, cleft palate, immune disorders, deafness and blindness. Thus, this project could illuminate the mechanisms of diverse disease states.

描述（由申请人提供）：无菌α基序（SAM）结构域是真核细胞中最常见的蛋白质模块之一。它们是在许多关键的调节蛋白，脚手架蛋白和转录因子中发现的，其中包括许多参与遗传疾病的蛋白质。对孤立系统的重点研究表明，SAM结构域是多功能蛋白质 - 蛋白质相互作用伴侣，形成同性恋者，异弱者，杂音异构体，同型聚合物，杂聚合物以及与其他大分子的复合物。然而，绝大多数SAM领域在功能上仍然没有表征，这在我们对重要的生物途径和疾病状态的理解中留下了重大差距。在这里，我们试图利用我们使用SAM领域结构和功能开发的大量专业知识来阐明SAM域在人类基因组中的功能。目的是：目标I：识别人类SAM聚合物。每个人类SAM结构域都将表示为增压GFP的融合，以保持聚合物溶解度。将通过天然凝胶和/或凝胶过滤色谱法筛选SAM域以进行骨料形成。如果发现它们形成聚集体，则将通过电子显微镜或AFM进一步检查它们，以将聚集体与真正的聚合物区分开。 AIM II：确定所有人类同型和杂种SAM相互作用都会发现聚合物调节剂和新共聚物。我们将使用拆分DHFR分析来识别彼此结合的SAM。确定的相互作用将通过AIMI的GFP融合来对生化进行验证。我们还希望发现新的SAM聚合物调节剂和可能的SAM共聚物。我们认为，这里提出的面向发现的方法是一种有效的方法，可以最大程度地提高我们先前的NIH资助的效用，从而使他人受益。 公共卫生相关性：细胞功能需要蛋白质之间的精确通信。当这些交流出现问题时，就会发生疾病。该项目旨在识别和表征大约一百个人蛋白质中发现的蛋白质结合模块的相互作用，包括直接参与自闭症，白血病，left裂，免疫疾病，耳聋和失明的蛋白质。因此，该项目可以阐明各种疾病状态的机制。