CHAIN GENE ORGANIZATION IN TRANSFORMED B LYMPHOCYTES

这项赠款及其前身的目标是研究 与重链基因表达相关的分子事件。 最后一 格兰特专注于C/H基因的结构和重排。 小鼠Mu-C δ基因座的深入研究 人类提供了一个框架来解决 转录和转录后调控的关键。 在这 更新，我们建议延长穆三角洲调节的工作， 主要是利用体外表达技术， 来理解基因产物的不同作用。 具体来说，我们将确定IgM和IgD的水平， 在人B细胞中受到调节;重要性来自于 在这一地区，C三角洲结构的巨大进化分歧 物种，同时保留IgD功能的表型保守性。 我们提出了一系列的转染研究，旨在罚款 3'的切割和鼠μ-δ基因座中的调节。 的 是什么机制（S）管理膜与分泌 羧基末端和/或Mu与δ外显子选择？ 我们提出 V/H启动子相关序列的研究，重点是 蛋白质：DNA相互作用和T细胞的调节作用 在这种互动上。 我们提出了一种新的cDNA克隆策略 用于表征推定的转录终止， 启动子相关因素。 我们提供初步的可行性 开发抗原特异性IgM的数据和设计方案 和IgD转染的B细胞系。 具体来说，我们将调查 磷酸胆碱特异性mIg受体的作用，无论是在野生型， 型和结构修饰形式，在B细胞活化中， 加工和抗原呈递。 最后，我们设计了一个 分子方法来解决的差异作用mIgM和 IgD在新生儿B细胞耐受中的作用

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.