ORGANIZATION AND EXPRESSION OF V GENES

V基因的组织和表达

• 批准号: 2178201
• 负责人: PETER H BRODEUR
• 金额: $ 21.22万
• 依托单位: TUFTS UNIVERSITY BOSTON
• 依托单位国家: 美国
• 财政年份: 1985
• 资助国家: 美国
• 起止时间: 1985-07-01 至 1998-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2178201
• 关键词: B lymphocyte; DNA footprinting; animal genetic material tag; antibody formation; cell differentiation; chromatin; gene expression; gene rearrangement; genetic regulatory element; genetic transcription; immunoglobulin genes; laboratory mouse; nucleic acid sequence; tissue /cell culture; transfection

This research project proposal is directed at the molecular and cellular mechanisms which control the qualitative and quantitative utilization of variable region genes during B cell development. The studies proposed are based on the premise that constraints in V gene use reflect the evolution of the rearrangement process to ensure the generation of a functional and protective pre-immune repertoire. Ongoing studies on V gene utilization focus on two well-characterized VK genes which are utilized at markedly different frequencies in BALB/c mice and for which there is evidence of rearrangement frequency differences, cellular selection, and intrinsic junctional bias. New studies are directed at identifying alterations of higher order chromatin structure within the Igh-V locus associated with VH gene rearrangement. These studies are based on the hypothesis that V gene rearrangement is tightly regulated by chromatin structure. Nuclease hypersensitivity studies will be used to determine the degree of VH gene exposure (accessibility) in different subregions throughout the Igh-V locus in non-rearranging and actively rearranging pre-B cell lines. Attempts will be made to identify a postulated cis-regulatory region responsible for regulating Igh-V locus rearrangement via VH exon accessibility. Other studies will identify regulatory elements associated with individual VH genes which function during V gene rearrangement. These studies on the molecular mechanisms and cellular mechanisms of antibody repertoire formation will provide insight into a process fundamental to the understanding of immune responsiveness. The available cell lines, molecular probes, and detailed mapping studies of the Igh locus provide a unique opportunity to examine the regulation of a large chromosomal region. These studies also impact directly on questions relating to V gene utilization biases of certain B cell neoplasms and the ability to reconstitute the antibody repertoire in bone marrow transplant patients.

该研究项目提案针对分子和细胞 控制定性和定量利用的机制 B细胞发育过程中的可变区基因。提议的研究是 基于V基因使用的限制反映进化的前提 的重新安排过程，以确保生成功能性和 保护性免疫前组库。 V基因利用的持续研究 重点关注两个特征明确的 VK 基因，它们的利用显着 BALB/c 小鼠的不同频率，有证据表明 重排频率差异、细胞选择和内在 连接偏向。新的研究旨在确定 与 VH 相关的 Igh-V 位点内的高级染色质结构 基因重排。这些研究基于 V 基因的假设 重排受到染色质结构的严格调控。核酸酶 超敏反应研究将用于确定VH基因的程度 整个 Igh-V 不同次区域的暴露（可及性） 非重排和主动重排前 B 细胞系中的位点。 将尝试确定假定的顺式监管区域 负责通过 VH 外显子调节 Igh-V 位点重排 可达性。其他研究将确定相关的监管要素 具有在V基因重排期间起作用的单个VH基因。 这些关于分子机制和细胞机制的研究 抗体库的形成将提供对过程的深入了解 是理解免疫反应的基础。可用的 Igh 的细胞系、分子探针和详细图谱研究 位点提供了一个独特的机会来检查大型的监管 染色体区域。这些研究也直接影响问题 与某些 B 细胞肿瘤的 V 基因利用偏差有关 在骨髓移植中重建抗体库的能力 患者。