Transcription regulation of B-Cells by TFII-I

TFII-I 对 B 细胞的转录调节

• 批准号: 7156804
• 负责人: Shauna R Hutchinson
• 金额: $ 3.81万
• 依托单位: TUFTS UNIVERSITY BOSTON
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-09-04 至 2009-09-03
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7156804
• 关键词: B lymphocyte; chromatin immunoprecipitation; cytogenetics; fluorescent in situ hybridization; functional /structural genomics; gel mobility shift assay; gene rearrangement; genetic promoter element; immunoglobulin genes; minority health professional; predoctoral investigator; protein binding; protein protein interaction; replicase; transcription factor; transfection /expression vector

DESCRIPTION (provided by applicant): TFII-I is a multi functional, signal dependent transcription factor that is implicated in the regulation of many genes. The purpose of this work is to determine its role in the immune system specifically in the Immunoglobulin heavy chain (IgH) locus of B-cells. It was previously shown in vitro that two TFII-I family members bound to a region of the IgH promoter named Downstream Immunological Control Element (DICE). However, these family members, TFII-I and BEN, have opposing transcriptional functions. Specific goals of the project include; determining how TFII-I and BEN transcription factors are recruited to the rearranged IgH promoter in vivo. This will be accomplished by the use of EMSA analysis with TFII-I and BEN mutant proteins, chromatin immunoprecipitation of the DICE region using TFII-I and BEN antibodies and functional assays to determine the potential role that these factors play. Because the IgH locus is also regulated at the level of subnuclear positioning, by virtue of their distinct function and subnuclear localization, TFII-I and BEN may be involved in this movement. If so, then 3D-FISH analysis of both knockdown and wild type B-cells might determine their potential role on the movement of the locus.

描述（由申请人提供）：TFII-I是一种多功能、信号依赖性转录因子，涉及许多基因的调节。 这项工作的目的是确定其在免疫系统中的作用，特别是在 B 细胞的免疫球蛋白重链 (IgH) 位点中的作用。 先前的体外研究表明，两个 TFII-I 家族成员与 IgH 启动子的一个名为下游免疫控制元件 (DICE) 的区域结合。 然而，这些家族成员 TFII-I 和 BEN 具有相反的转录功能。 该项目的具体目标包括：确定 TFII-I 和 BEN 转录因子如何在体内招募至重排的 IgH 启动子。 这将通过使用 TFII-I 和 BEN 突变蛋白的 EMSA 分析、使用 TFII-I 和 BEN 抗体对 DICE 区域进行染色质免疫沉淀以及功能测定来确定这些因素发挥的潜在作用来完成。 由于 IgH 位点也在亚核定位水平上受到调控，凭借其独特的功能和亚核定位，TFII-I 和 BEN 可能参与了这场运动。 如果是这样，那么对敲低型和野生型 B 细胞进行 3D-FISH 分析可能会确定它们在基因座运动中的潜在作用。