Transcription regulation of B-Cells by TFII-I

描述(申请人提供)：TFII-I是一种多功能的、信号依赖的转录因子，参与许多基因的调节。这项工作的目的是确定它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)区域。先前在体外研究表明，两个TFII-I家族成员与IgH启动子的下游免疫控制元件(DICE)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：确定在体内如何将TFII-I和BEN转录因子招募到重排的IgH启动子。这将通过对TFII-I和BEN突变蛋白进行EMSA分析、使用TFII-I和BEN抗体对骰子区域进行染色质免疫沉淀以及功能分析来确定这些因子所起的潜在作用来实现。由于免疫球蛋白基因位点也在亚核定位水平上受到调控，由于其独特的功能和亚核定位，TFII-I和BEN 可能参与了这场运动。如果是这样的话，那么对敲除和野生型B细胞的3D-FISH分析可能会确定它们在基因座移动中的潜在作用。

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.