CELLULAR LOCALISATION OF THE TRANSCRIPTIONAL REGULATORS E12 AND E47

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 E2a蛋白E12和E47在B细胞发育过程中表达，以激活B细胞特异性基因表达和免疫球蛋白基因重排。在这里，我们建议使用二砷-四半胱氨酸标记来检查B细胞发育不同阶段E12和E47的细胞定位。E12和E47是转录调节因子，控制发育中的淋巴样细胞的发育进程、细胞扩张、存活和基因重排。E12和E47如何调控这一系列不同的生物活动尚不清楚。此外，在淋巴细胞发育的不同阶段，E12和E47的定位尚不清楚。在本申请中，我们建议使用双砷-四半胱氨酸表位来标记E12和E47。我们将通过荧光和EM研究来检查定位。最终，我们希望结合电子显微镜和3D-FISH来检查这些转录因子在免疫球蛋白基因座上的定位。从这些研究中获得的数据可能会对免疫球蛋白基因重排的单等位基因激活有深入的了解。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The E2A proteins E12 and E47 are expressed throughout B-cell development to activate B-cell specific gene expression and immunoglobulin gene rearrangement. Here we propose to examine the cellular localization of E12 and E47 during distinct stages of B-cell development utilizing biarsenical-tetracysteine labeling. E12 and E47 are transcriptional regulators that control developmental progression, cellular expansion, survival and gene rearrangement in developing lymphoid cells. How E12 and E47 regulate this diverse set of biological activities is unknown. Additionally it is unclear where E12 and E47 are localized during the distinct stages of lymphocyte development. In this application we propose to tag E12 and E47 utilizing the biarsenical-tetracysteine epitope. We would examine localization by fluorescence as well as EM studies. Ultimately we would like to combine electron microscopy and 3D-FISH to examine the localization of these transcription factors in the immunoglobulin locus. Data obtains from the studies may obtain insights into the mono-allelic activation of immunoglobulin gene rearrangement.