CHARACTERIZATION OF AN ETS RELATED TRANSCRIPTION FACTOR

描述：（改编自申请人的摘要） 该项目是一种新的转录因子PU. 1的表征， 在巨噬细胞和B细胞中表达。 PU. 1的DNA结合位点是 富含嘌呤的序列，5 ′-GAGGAA-3 ′（PU框）。 PU. 1是一个转录的 可能是基因表达的重要调节因子， 巨噬细胞和B细胞。 PU.1的羧基末端氨基酸序列 与氨基酸序列的一个区域具有显著的同一性（41%）， 在ETS致癌基因家族中。 这个区域包含了 1的DNA结合结构域，可能是ets的DNA结合结构域 proteins. 第一个具体目标是表征DNA结合结构域， PU.1的推定激活结构域。通过缺失分析和体外 诱变 将详细定义这些区域。 的信息 从这项研究，希望能提供一个更好的了解如何PU.l 蛋白质与DNA相互作用并能够激活转录。 第二个具体目标是研究PU.实验 已被设计用于分离可能受PU.1调节的基因。一些 将评估含有PU. 1结合位点的基因启动子， 将确定PU.l结合位点是否需要 转录。 识别PU. 1的功能的其他方法包括 有义和反义PU.1的过量表达。的 PU. 1的调节可对细胞生长或活化具有显著影响。 将在巨噬细胞和B细胞中检测PU. 1蛋白的水平 用细胞因子或其它活化剂处理。 这些结果可能与 PU. 1具有巨噬细胞和B细胞的特异性功能。 本项目的长期目标是更好地了解 免疫系统中的转录调节。 细胞类型特异性 转录因子PU.1可能提供了实现这一点的机会 objective.

DESCRIPTION: (Adapted from the applicant's abstract) The focus of this project is the characterization of a new transcription factor, PU.l which is expressed in macrophages and B cells. The DNA binding site for PU.l is a purine rich sequence, 5' -GAGGAA-3' (PU box). PU.l is a transcriptional activator that may be an important regulator of gene expression in macrophages and B cells. The carboxy terminal amino acid sequence of PU.1 has significant identity (41 percent) with a region of amino acid sequence in the ets oncogene family. This region of identity contains the DNA binding domain of PU.l and is probably the DNA binding domain of ets proteins. The first specific aim is to characterize the DNA binding domain and putative activation domain of PU.l. By deletion analysis and in vitro mutagenesis. These regions will be defined in detail. The information from this study is hoped to provide a better understanding of how the PU.l protein interacts with DNA and is able to activate transcription. The second specific aim is to study the function of PU.l. Experiments have been designed to isolate genes that may be regulated by PU.l. A number of gene promoters which contain PU.l binding sites will be evaluated and a determination will be made as to whether PU.l binding site is necessary for transcription. Other approaches to identify the function of PU.l include the over-expression of sense and antisense constructions of PU.l. The modulation of PU.l may have dramatic effects on cell growth or activation. The level of PU.l protein will be examined in macrophages and B cells treated with cytokines or other activating agents. These results may link PU.l with specific functions of macrophages and B cells. The long-term objectives of this project are to gain a better understanding of transcriptional regulation in the immune system. The cell type specific transcription factor PU.l may provide an opportunity to achieve this objective.