CHARACTERIZATION OF AN ETS RELATED TRANSCRIPTION FACTOR

描述：(改编自申请者的摘要)本次活动的重点 项目是对一种新的转录因子PU.l的鉴定 在巨噬细胞和B细胞中表达。PU.l的DNA结合部位是 富嘌呤序列，5‘-GAGGAA-3’(PU盒)。PU.l是一种转录 激活因子可能是一种重要的基因表达调控因子 巨噬细胞和B细胞。PU1的羧基末端氨基酸序列 与氨基酸序列的一个区域有显著的同源性(41%) 在ETS癌基因家族中。这个身份区域包含DNA PU的结合域，可能是Ets的DNA结合域 蛋白质。 第一个具体目标是表征DNA结合域和 PU的可能激活结构域。通过缺失分析和体外实验 诱变。将对这些区域进行详细定义。这些信息 通过这项研究，希望能更好地了解PU是如何 蛋白质与DNA相互作用，能够激活转录。 第二个具体目的是研究PU.l的功能。实验已经 被设计用来分离可能受PU.l调控的基因。一批 将对包含PU结合位点的基因启动子进行评估，并 将确定PU.l结合位点是否为 抄写。识别PU功能的其他方法包括 PU的正反义结构的过度表达这个 PU的调节可能会对细胞的生长或激活产生显著的影响。 将在巨噬细胞和B细胞中检测PU.l蛋白的水平 用细胞因子或其他激活剂治疗的。这些结果可能会将 PU.l具有巨噬细胞和B细胞的特殊功能。 这个项目的长期目标是为了更好地了解 在免疫系统中的转录调控。单元格类型特定 转录因子PU.1可能为实现这一目标提供了机会 目标。

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.