Post-transcriptional Control of Immunoglobulin Expression

免疫球蛋白表达的转录后控制

• 批准号: 9507513
• 负责人: Martha Peterson
• 金额: $ 30万
• 依托单位: University of Kentucky Research Foundation
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 1995
• 资助国家: 美国
• 起止时间: 1995-08-15 至 1999-07-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=9507513&HistoricalAwards=false
• 关键词: Post; transcriptional; Control; Immunoglobulin; Expression

9507513 Peterson Regulated transcriptional termination and RNA processing of the immunoglobulin (Ig) heavy chain ( gene will be examined during B cell development. The long-term goal of this research is to better understand the general mechanisms governing these events as well as to determine the basis for the developmentally regulated changes that occur during B cell maturation. The regulated alternative processing of a ( precursor RNA to form (s and (m mRNA does not require any Ig gene-specific sequences. Instead, the important feature of this gene is that it contains signals for competing splice and cleavage-polyadenylation reactions. Thus, the amount or activity of a general factor(s) involved in these two processes must be altered during B cell maturation. It will be determined whether splicing is regulated during B cell maturation by analyzing alternative splice choices made in the absence of a poly(A) site in B cells as compared to plasma cells. An in vitro splicing/polyadenylation system that will mimic in vivo ( gene characteristics will be examined. Successfully establishing an in vitro system to study ( regulation would provide the basis for many future experiments. The expression of the known general RNA processing regulators in B cells and plasma cells will be examined to identify factors that potentially mediate the regulation; any whose expression correlates with ( processing changes will be studied further by over-expression in B cells and/or plasma cells. These experiments, taken together, will provide a more complete picture of ( processing regulation and should begin to identify the trans-acting RNA processing components that are altered during B cell maturation. To investigate transcriptional termination and its regulation, the transcription of intact and modified ( genes, stably introduced into plasma cell and B cell lines, will be analyzed by nuclear run-on experiments to identify their termination regions. Gene modifications will be designed to determine the contributio n that the two ( poly(A) sites, the poly(A) site strength, and the downstream sequences within the termination region make to the overall termination process. This study, by comparing normal and modified genes in a single cell type will provide information about the general process of termination and, by comparing the modified genes in both B cells and plasma cells, will examine the basis for regulatory changes in this process. %%% Regulated transcriptional termination and RNA processing of the immunoglobulin (Ig) heavy chain ( gene will be examined during B cell development. The long-term goal of this research is to better understand the general mechanisms governing these events as well as to determine the basis for the developmentally regulated changes that occur during B cell maturation. ***

9507513彼得森调控的免疫球蛋白(Ig)重链(基因)转录终止和核糖核酸加工将在B细胞发育过程中进行检测。这项研究的长期目标是更好地了解控制这些事件的一般机制，以及确定在B细胞成熟过程中发生的发育调节变化的基础。A(前体RNA的受调控的交替加工形成(S和(M)mRNA不需要任何Ig基因特异性序列。相反，该基因的重要特征是它包含竞争剪接和切割-聚腺苷酸化反应的信号。因此，在B细胞成熟过程中，参与这两个过程的一种通用因子(S)的数量或活性必须改变。这将通过分析在B细胞中没有Poly(A)位点的情况下与浆细胞相比所做出的选择性剪接选择来确定剪接是否在B细胞成熟过程中受到调控。一种体外剪接/聚腺苷酸化系统，它将模仿体内(基因特征将被检测。成功地建立了一个体外系统来研究(调控将为未来的许多实验提供基础。已知的通用RNA加工调节因子在B细胞和浆细胞中的表达将被检测，以确定可能介导调节的因素；任何其表达与(加工变化)相关的因素将通过在B细胞和/或浆细胞中的过度表达来进一步研究。这些实验结合在一起，将提供一个更完整的加工调控图景，并应该开始识别在B细胞成熟过程中改变的反式作用RNA加工成分。为了研究转录终止及其调控，稳定地导入浆细胞和B细胞系的完整和修饰基因的转录将通过核连续实验进行分析，以确定它们的终止区。基因修饰将被设计成确定两个聚(A)位点、聚(A)位点强度和终止区内的下游序列对整个终止过程的贡献。这项研究通过比较单个细胞类型中的正常基因和修饰基因，将提供有关终止的一般过程的信息，并通过比较B细胞和浆细胞中的修饰基因，将检验这一过程中调控变化的基础。免疫球蛋白(Ig)重链基因的转录终止和RNA加工将在B细胞发育过程中进行检查。这项研究的长期目标是更好地了解控制这些事件的一般机制，以及确定在B细胞成熟过程中发生的发育调节变化的基础。***