Regulation of V(D)J recombination by Rag2 C terminus

Rag2 C 末端对 V(D)J 重组的调节

基本信息

批准号：
8260320
负责人：
PATRICIA CORTES
金额：
$ 33.56万
依托单位：
ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-05-20 至 2015-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8260320
关键词：
Accounting Acidic Region Acids Address Affect Amino Acids Antibodies Applications Grants B-Lymphocytes Binding Biochemical Biochemistry C-terminal Cell Surface Receptors Cell Survival Chromatin Complex DNA DNA Binding DNA biosynthesis Development Fingers Generations Genes Genetic Recombination Goals Histone H3 Histones Immune system Immunoglobulins Immunologic Deficiency Syndromes J segment gene Lead Length Lymphocyte Lysine MCM2 gene MCM4 gene Mediating Modification Molecular Mutation Nucleosomes Patients Plants Play Principal Investigator Process Proteins Rag1 Mouse Reaction Regulation Reporting Role Series Site Solutions Stretching T-Cell Receptor T-Lymphocyte V(D)J Recombination Work chromatin immunoprecipitation homeodomain in vivo leukemia/lymphoma mutant pathogen programs protein protein interaction protocol development public health relevance recombinase repaired research study

项目摘要

DESCRIPTION (provided by applicant): V(D)J recombination is a site-specific somatic recombination reaction whose end products are genes encoding cell surface receptors and antibody molecules that are central to immune system function. Proper recombination is absolutely required for development of a healthy immune system: when the reaction does not occur or is poorly controlled, there is the potential for development of immunodeficiencies, leukemias, and lymphomas. Our long-term goal is to understand the mechanisms of V(D)J recombination and its regulation, and our approach has been through a biochemical analysis of the RAG proteins. Though much progress has been made in the field of recombination in the last two decades, there are still numerous questions outstanding, among them are the issue of accessibility and the regulation of recombination in vivo. In this grant application, we describe a series of experiments that will address these issues while extending our findings that showed interaction of Rag2 with core histones and the MCM complex. Through specific aims proposed, we will: 1) Investigate the requirement and participation of MCM2-7 complex and MCM associated proteins in V(D)J recombination; 2) Study the role of Rag2 C-terminus, acidic region and PHD domain, in the different steps of V(D)J recombination as well as their participation in the various protein-protein interactions mediated by this region; 3) Analyze the RSS cleavage mechanisms that depend on Rag2 full-length using a chromatin substrate. The focus of these aims is to understand the molecular mechanism that determine regulation of V(D)J recombination by Rag2 and specifically the contribution of Rag2 C-terminus to this process. PUBLIC HEALTH RELEVANCE: V(D)J recombination is an essential process that takes place in B and T lymphocytes. This process is mediated by recombination factors that play unique and fundamental roles; deregulation of any of the required factors results in severe immunodeficiency. This project is focused on understanding how Rag2, one of the recombination factors, regulated the reaction by mediating multiple protein interactions and thus modulating the generation of a diverse immune system.

描述（由申请人提供）：V（d）J重组是一种特定位点的体细胞重组反应，其最终产物是编码细胞表面受体和抗体分子的基因，对免疫系统功能核心。发展健康免疫系统绝对需要适当的重组：当反应不发生或控制不善时，可能会发展免疫缺陷，白血病和淋巴瘤。我们的长期目标是了解V（d）J重组及其调节的机制，我们的方法是通过对抹布蛋白的生化分析。尽管在过去的二十年中，在重组领域取得了很多进展，但仍有许多问题，其中包括可访问性和体内重组的调节问题。在此赠款应用程序中，我们描述了一系列实验，这些实验将解决这些问题，同时扩展了我们的发现，这些发现显示了RAG2与核心组蛋白和MCM复合物的相互作用。通过提出的特定目的，我们将：1）研究MCM2-7复合物和MCM相关蛋白在V（d）J重组中的需求和参与； 2）研究RAG2 C末端，酸性区域和PHD结构域的作用，在V（d）J重组的不同步骤中以及它们参与由该区域介导的各种蛋白质蛋白质相互作用； 3）分析使用染色质底物依赖RAG2全长的RSS裂解机制。这些目的的重点是了解确定RAG2的V（d）J重组调节的分子机制，特别是RAG2 C端对此过程的贡献。公共卫生相关性：V（d）J重组是在B和T淋巴细胞中发生的重要过程。此过程是由重组因素介导的，这些因素起着独特而基本的作用。对任何所需因素的放松管制会导致严重的免疫缺陷。该项目的重点是理解RAG2是如何通过介导多种蛋白质相互作用并从而调节多种免疫系统的产生来调节反应的一种。