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

E2A蛋白作为早期B细胞形成的有效调节剂的重要性已得到证实，但其在成熟B细胞向浆细胞转变中的作用尚不清楚。最近，一种新的含有e2a的E-box结合物质，被称为P-E2A，在成熟的B-向浆细胞转变的过程中被检测到。本研究的目的包括：1)研究阶段特异性e2a复合物P-E2A的转录成分和作用；2)鉴定浆细胞中与E2A相互作用的其他蛋白；3)表征这些e2a相互作用蛋白。为了实现这些特定的目标，蛋白质-蛋白质相互作用克隆系统将被用来描述P-E2A和其他e2a相互作用蛋白复合物的分子成分，这些蛋白复合物是浆细胞所特有的。编码潜在的e2a相互作用蛋白的基因的检索将随后使用广泛的遗传和生化技术进行分析，包括基因克隆和测序，RNA和免疫印迹，电泳迁移分析，以及在各种培养细胞中的DNA转染。其他局限于与浆细胞不同的细胞类型的e2a相互作用蛋白已被鉴定并证明是细胞分化和增殖等过程的重要转录调节因子。该项目将确定在浆细胞中发现的e2a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（如抗体基因转录）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控制细胞限制性真核基因表达的分子机制的数据。E2A蛋白是免疫系统的关键转录调节因子。特别是，E2A基因产物是合成抗体的特化细胞发育所必需的。该项目将增强对E2A相关蛋白的认识，并深入了解E2A与各种调节蛋白之间的相互作用如何导致dna -蛋白复合物的形成，从而调节浆细胞中抗体的产生。

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.