Post-transcriptional Control of Immunoglobulin Expression

智力价值：这项研究的长期目标是了解基因表达的调节，特别是关注基因转录和RNA加工之间的联系。虽然关于这些过程的各个方面已经积累了大量的知识，但它们是如何相互联系的，以及这种相互作用如何影响监管还有待确定。该项目将探索RNA加工事件(剪接和切割-多聚腺苷基化)与免疫球蛋白M(IgM或µ)基因的转录延长和终止之间的机械相互联系。µ基因是解决这些问题的一个很好的模型系统，因为它在B淋巴细胞成熟过程中的表达受到多个步骤的调节，包括RNA剪接、切割-聚腺苷酸化和转录终止。对于这项工作，PI的实验室提供了大量具有良好特性的修改后的µ基因、用于分析µ基因表达的多种试剂以及在刺激分化前后B细胞的微阵列数据。该项目将基于微阵列数据和基因表达实验，确定µ替代RNA加工的反式调节因子，结合核运行和高分辨率染色质免疫沉淀(CHIP)分析，确定RNA加工信号如何影响RNA聚合酶II的伸长，并通过高分辨率芯片实验表征与延长B细胞和浆细胞中野生型和修饰的µ基因相关的因素的变化。这些研究将有助于更好地理解µ基因中转录和RNA加工之间的联系，并将为这些联系提供新的线索，这些联系也调节许多其他基因的表达。广泛影响这项研究的更广泛影响包括研究生、本科生和可能的高中生参与研究，以及他们参加地方和国家科学会议。在过去的18年里，与PI的实验室相关的大多数学生都是女性。与该实验室的一名前研究生，目前是中田纳西州州立大学副教授的持续合作将继续进行；微阵列数据一直是本科生实验室课程和个别学生独立研究项目的基础。这笔赠款的PI继续以科学博览会评委的身份参与并访问高中课堂，以支持更广泛的社区的科学。此外，作为肯塔基大学负责研究的兼职副校长和KY科学院成员，PI支持肯塔基全州机构的研究。该项目产生的数据和资源将通过国家和国际会议介绍和出版物与科学界广泛共享。

Intellectual Merit:The long-term goal of this research is to understand the regulation of gene expression, focusing specifically on the connections between gene transcription and RNA processing. While a substantial base of knowledge has been accumulated about these processes individually, how they are coupled and how this coupling affects regulation has yet to be determined. This project will explore the mechanistic interconnections between RNA processing events (splicing and cleavage-polyadenylation) and transcription elongation and termination in the immunoglobulin M (IgM or µ) gene. The µ gene is an excellent model system in which to address these questions since its expression is regulated during B lymphocyte maturation at multiple steps, including RNA splicing, cleavage-polyadenylation and transcription termination. An extensive collection of well-characterized modified µ genes, multiple reagents to analyze µ gene expression and microarray data from B cells before and after being stimulated to differentiate, are available in the PI's lab for this work. The project will identify trans-regulators of µ alternative RNA processing based on microarray data and gene expression experiments, determine how RNA processing signals affect RNA polymerase II elongation, using a combination of nuclear run-on and high-resolution chromatin immunoprecipitation (ChIP) assays and characterize changes in factors associated with elongating pol II over wild-type and modified µ genes in B cells and plasma cells by high-resolution ChIP experiments. These studies will lead to a better understanding of the connections between transcription and RNA processing in the µ gene and will shed new light on these connections, which also regulate the expression of many other genes.Broader ImpactsThe broader impacts of this research include the involvement of graduate, undergraduate, and possibly high school students in the research, as well as their participation in local and national scientific meetings. The majority of students associated with the PI's lab over the past eighteen years have been female. An on-going collaboration with a former graduate student from this lab, who is currently an Associate Professor at Middle TN State University, will continue; the microarray data has been the basis for projects in undergraduate lab courses and individual student independent research projects. The PI of this grant continues to support science in the broader community by participating as a science fair judge and visiting high school classrooms. In addition, as a part-time Associate Vice President for Research at the University of Kentucky and member of the KY Academy of Sciences, the PI supports research in institutions across the state of KY. The data and resources generated from this project will be shared widely with the scientific community through national and international meeting presentations and publications.