Transcription regulation of B-Cells by TFII-I

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。先前的体外研究表明，两个TFII-I家族成员与IgH启动子的下游免疫控制元件（DICE）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。项目的具体目标包括：确定TFII-I和BEN转录因子如何在体内被募集到重排的IgH启动子上。这将通过使用TFII-I和BEN突变蛋白进行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所起的潜在作用来完成。由于IgH位点也在亚核定位水平受到调控，凭借其不同的功能和亚核定位，TFII-I和BEN

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.