Transcription regulation of B-Cells by TFII-I

TFII-I 对 B 细胞的转录调节

• 批准号: 7928012
• 负责人: Shauna R Hutchinson
• 金额: $ 3.83万
• 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 2010-09-03
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7928012
• 关键词: Antibodies; B-Cell Development; B-Lymphocytes; Binding; Biochemical; Biological Assay; Cells; Chromatin; DNA-Binding Proteins; Data; Down-Regulation; Electrophoretic Mobility Shift Assay; Elements; Excision; Family; Family member; Fellowship; Fluorescent in Situ Hybridization; Genes; Goals; Heavy-Chain Immunoglobulins; Human; IGH@ gene cluster; Immune system; In Vitro; Individual; Mediating; Movement; Mus; Names; Nuclear; Phenotype; Play; Positioning Attribute; Precipitation; Proteins; Recruitment Activity; Regulation; Role; Signal Transduction; Transcriptional Activation; Transcriptional Regulation; Transfection; Work; chromatin immunoprecipitation; in vivo; knock-down; mutant; promoter; protein complex; research study; transcription factor

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，与许多基因的调节有关。 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）基因座中。 以前在体外显示，两个TFII-I家族成员与名为下游免疫控制元件（DICE）的IGH启动子的区域结合。 但是，这些家庭成员TFII-I和Ben具有相反的转录功能。 该项目的具体目标包括：确定如何将TFII-I和BEN转录因子募集到体内重排的IGH启动子。 这将通过使用TFII-I和BEN突变蛋白，使用TFII-I和BEN抗体和功能测定法对骰子区域的染色质免疫沉淀来确定这些因素的潜在作用。 由于IGH基因座也受到亚核定位水平的调节，由于其独特的功能和亚核定位，因此 可能参与了这一运动。 如果是这样，那么对敲低和野生型B细胞的3D-FISH分析可能会决定它们在基因座运动中的潜在作用。