SAM Domains

SAM 域

• 批准号: 8245084
• 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/8245084
• 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 Alpha 基序 (SAM) 结构域是真核细胞中最常见的蛋白质模块之一。它们存在于许多关键调节蛋白、支架蛋白和转录因子中，其中包括许多与遗传疾病有关的蛋白质。对孤立系统的集中研究表明，SAM 结构域是多功能的蛋白质-蛋白质相互作用伙伴，可形成同源寡聚物、异源寡聚物、同聚物、异聚物以及与其他大分子的复合物。然而，绝大多数 SAM 结构域在功能上仍然没有特征，这使得我们对重要生物途径和疾病状态的理解存在重大差距。在这里，我们寻求利用我们在 SAM 结构域结构和功能方面积累的大量专业知识，从全球角度阐明人类基因组中 SAM 结构域的功能。目标是： 目标 I：识别人类 SAM 聚合物。每个人-SAM 结构域都将表达为与增压 GFP 的融合体，以维持聚合物的溶解度。将通过天然凝胶和/或凝胶过滤色谱法筛选 SAM 结构域的聚集体形成。如果发现它们形成聚集体，将通过电子显微镜或 AFM 进一步检查，以区分聚集体和真正的聚合物。目标 II：识别所有人类同源和异源 SAM 相互作用，发现聚合物调节剂和新共聚物。我们将使用分离 DHFR 测定法来识别彼此结合的 SAM。已确定的相互作用将通过 Aim I 的 GFP 融合体进行生化验证。我们还希望发现新的 SAM 聚合物调节剂以及可能的 SAM 共聚物。我们相信，这里提出的以发现为导向的方法是一种有效的方法，可以最大限度地利用我们之前 NIH 资助的进展，造福他人。 公共健康相关性：细胞功能需要蛋白质之间的精确通信。当这些交换出现问题时，疾病就会发生。该项目旨在识别和表征大约一百种人类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、腭裂、免疫疾病、耳聋和失明的蛋白质。因此，该项目可以阐明不同疾病状态的机制。