CHAIN GENE ORGANIZATION IN TRANSFORMED B LYMPHOCYTES

这项拨款及其前身的目的是研究 与重链基因表达相关的分子事件。最后 格兰特主要研究C/H基因的结构和重排。 小鼠Mu-C增量基因座的深入研究 人类为解决根本问题提供了一个框架 转录和转录后调控。在这 更新，我们建议延长Mu-Delta规则的工作， 主要是开发体外表达技术，用眼睛 了解基因产物的不同作用。 具体地说，我们将确定IgM和IGD的水平 在人类B细胞中被调节；意义来自于 本区C三角洲构造演化差异显著 在IGD功能中保留表型保守性的物种。 我们提出了一系列旨在改善 小鼠Mu-Delta基因3‘端的解剖与调控。那 是，膜与分泌的调控机制(S)是什么 羧基末端和/或Mu与Delta外显子选择？我们建议 V/H启动子相关序列的研究，重点是 蛋白质：DNA相互作用与T细胞的调节作用 在这种互动中。我们提出了一种新的cDNA克隆策略 用于表征假定的转录终止和 启动子相关因子。我们提供了初步的可行性 开发抗原特异性IgM的数据和设计方案 并将IGD基因导入B细胞系。具体地说，我们将调查 磷酸胆碱特异性MIG受体的作用，无论是在野生- 类型和结构修饰形式，在B细胞激活中， 处理和抗原呈递。最后，我们设计了一个 用分子方法研究mIgM和mIgM的不同作用 新生儿B细胞耐受中的IGD

The goal of this grant and its predecessor is to study the molecular events related to heave chain gene expression. The last grant focused on the structure and rearrangement of C/H genes. An in depth characterization of the Mu-C delta locus in mouse and man has provided a framework to address fundamental issues of transcriptional and post-transcriptional regulation. In this renewal, we propose to extend the work on Mu-delta regulation, primarily be exploiting in vitro expression technology, with an eye toward understanding the differential roles of the gene products. Specifically, we will determine the levels at which IgM and IgD are regulated in human B cells; significance draws on the dramatic evolutionary divergence of C delta structure in this species while retaining phenotypic conservation in IgD function. We propose a series of transfection studies aimed at fine dissection of 3' and regulation in the murine mu-delta locus. That is, what is the mechanism(s) governing membrane vs. secreted carboxyl-terminus and/or Mu vs delta exon selection? We propose studies on V/H promoter-associated sequences, focusing on protein: DNA interactions and the regulatory effects of T cells on this interaction. We propose a novel cDNA cloning strategy for characterizing putative transcription termination and promoter-associated factors. We provide preliminary feasibility data and design projects for exploitation of antigen-specific IgM and IgD transfected B cell lines. Specifically, we will investigate the role of phosphorylcholine-specific mIg receptors, both in wild- type and structurally modified forms, in B cell activation, processing, and antigen presentation. Finally, we devise a molecular approach to address the differential roles of mIgM and IgD in neonatal B cell tolerance.