RUI: The Isolation and Characterization of Basic Helix-Loop-Helix Proteins Associated with E2A in Plasma Cells

RUI：浆细胞中与 E2A 相关的碱性螺旋-环-螺旋蛋白的分离和表征

• 批准号: 9728774
• 负责人: Craig Vierra
• 金额: $ 12.86万
• 依托单位: University of the Pacific
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-10-01 至 2001-09-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=9728774&HistoricalAwards=false
• 关键词: RUI; Isolation; Characterization; Basic; Helix

Vierra The importance of E2A proteins as potent regulators of early B cell formation is well established, but their role in the mature B- to plasma cell transition remains unclear. Recently, a new E2A-containing E-box binding species, designated P-E2A, has been detected during the mature B- to plasma cell transition. The objectives of the research include 1) investigating the transcriptional components and role of the stage-specific E2A-containing complex, P-E2A; 2) identifying other protein(s) that interact with E2A in plasma cells; 3) characterizing these E2A-interacting protein(s). To accomplish these specific goals, protein-protein interactional cloning systems will be performed to delineate the molecular components of P-E2A and other E2A-interacting protein complexes unique to plasma cells. The retrieval of genes encoding potential E2A-interacting proteins will be subsequently analyzed using a broad spectrum of genetic and biochemical techniques, including gene cloning and sequencing, RNA- and immunoblotting, electrophoretic-mobility shift analyses, and DNA transfections in a variety of cultured cells. Other E2A-interacting proteins restricted to cell types distinct from plasma cells have been identified and demonstrated to be important transcriptional regulators of processes such as cellular differentiation and proliferation. This project will determine whether E2A-interacting transcription factors found in plasma cells play a similar role in processes such as plasma cell formation or cell-specific gene expression e.g. antibody gene transcription. The results of these studies will identify transcription factors whose pattern of expression is restricted to the plasma cell lineage and will provide data that will help to elucidate the molecular mechanisms that govern cell-restricted eukaryotic gene expression. E2A proteins are critical transcriptional regulators of the immune system. In particular, E2A gene products are required for the development of specialized cells that synthesize antibodies. This project will enhance the knowledge of E2A-associating proteins and provide insight into how interactions between E2A and various regulatory proteins result in the formation of DNA-protein complexes that regulate antibody production in plasma cells.

E2 A蛋白作为早期B细胞形成的有效调节剂的重要性已得到充分证实，但其在成熟B-浆细胞转化中的作用仍不清楚。 最近，在成熟的B细胞向浆细胞的转变过程中检测到一种新的含E2 A的E-box结合物质，命名为P-E2 A。本研究的目的包括：1）研究阶段特异性含E2 A复合物P-E2 A的转录组分和作用; 2）鉴定浆细胞中与E2 A相互作用的其他蛋白; 3）表征这些E2 A相互作用蛋白。 为了实现这些特定的目标，将进行蛋白质-蛋白质互作克隆系统，以描绘P-E2 A和浆细胞特有的其他E2 A相互作用蛋白复合物的分子组分。 随后将使用广泛的遗传和生物化学技术，包括基因克隆和测序、RNA和免疫印迹、细胞迁移率变化分析和各种培养细胞中的DNA转染，分析编码潜在E2 A相互作用蛋白的基因的检索。 其他E2 A相互作用蛋白质限制于细胞类型不同于浆细胞已被确定，并证明是重要的转录调控过程，如细胞分化和增殖。 该项目将确定浆细胞中发现的E2 A相互作用转录因子是否在浆细胞形成或细胞特异性基因表达（例如抗体基因转录）等过程中发挥类似作用。 这些研究的结果将确定其表达模式仅限于浆细胞谱系的转录因子，并将提供有助于阐明细胞限制性真核基因表达的分子机制的数据。 E2 A蛋白是免疫系统的关键转录调节因子。 特别是，E2 A基因产物是合成抗体的特化细胞发育所必需的。 该项目将增强E2 A相关蛋白的知识，并深入了解E2 A与各种调节蛋白之间的相互作用如何导致形成调节浆细胞中抗体产生的DNA-蛋白质复合物。