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

Rag2 C 末端对 V(D)J 重组的调节

• 批准号: 8458581
• 负责人: PATRICIA CORTES
• 金额: $ 31.55万
• 依托单位: ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
• 依托单位国家: 美国
• 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/8458581
• 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

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.

描述（由申请人提供）：V(D)J重组是一种位点特异性体细胞重组反应，其最终产物是编码对免疫系统功能至关重要的细胞表面受体和抗体分子的基因。正确的重组对于健康免疫系统的发育是绝对必要的：当反应不发生或控制不佳时，就有可能发展成免疫缺陷、白血病和淋巴瘤。我们的长期目标是了解 V(D)J 重组的机制及其调控，我们的方法是通过对 RAG 蛋白进行生化分析。尽管近二十年来重组领域取得了很大进展，但仍然存在许多悬而未决的问题，其中包括体内重组的可及性和调控问题。在本次拨款申请中，我们描述了一系列实验，这些实验将解决这些问题，同时扩展我们的发现，显示 Rag2 与核心组蛋白和 MCM 复合物的相互作用。通过提出的具体目标，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端、酸性区和PHD结构域在V(D)J重组不同步骤中的作用以及它们参与该区域介导的各种蛋白质-蛋白质相互作用； 3) 使用染色质底物分析依赖于 Rag2 全长的 RSS 切割机制。这些目标的重点是了解决定 Rag2 对 V(D)J 重组调节的分子机制，特别是 Rag2 C 末端对此过程的贡献。