BIOLOGICAL FUNCTION OF BASONUCLIN

描述：(改编自申请者的摘要)-长期目标 这一提议的关键是理解 维持细胞的增殖能力和触发的信号 细胞停止增殖。首席调查员认为 理解转录调控在从 增殖到分化是阐明肿瘤的重要一步 这些监管程序。首席调查员最近 发现了一种新的锌指蛋白，Basonuclin。巴索努克林在场 在表皮增殖区角质形成细胞的核中 以及睾丸生精小管中的生殖细胞。校长 研究人员假设Basonuclin是一种转录因子 参与细胞增殖潜能的调节 (能力)。具体目的1是证明Basonuclin是一种DNA 具有序列特异性的结合蛋白。方法是选择这些 用电泳法从随机序列池中提取靶点 使用锌指多肽进行检测(EMSA)。具体目标2将确定 启动子包含Basonuclin靶序列的基因。校长 研究人员致力于识别受碱性核蛋白控制的基因 从而了解basonuclin是一种 一部份。首席调查员将在DNA数据库中搜索 含有Basonuclin靶序列的启动子，并分析 选择基因组片段是因为它们有能力结合到 细菌产生全长的碱性核蛋白。《特定目标3》将绘制 有丝分裂染色体上的碱性核蛋白结合部位。联谊会 具有有丝分裂染色质的碱性核蛋白是非常罕见的。校长 研究人员的目标是识别包含有丝分裂的染色体 并克隆这些结合位点，这可能是也可能不是 裸露的DNA上存在相同的结合位点(缺失组蛋白)，即 研究目的2.碱性核蛋白等分子在研究中的优点 对细胞活动的生化和遗传调控是一种 也受到监管的监管机构。通过研究它，校长 研究人员希望不仅获得关于Basonuclin的知识，而且还 关于它是其中一部分的监管等级。校长 研究人员认为，拟议中的对脑血管内皮细胞生物学功能的研究 Basonuclin将增强对细胞周期蛋白分子调控的认识 细胞增殖潜能。他认为，这种知识是必要的，以 设计更好的医疗方法来治疗与扩散相关的异常 皮肤，如癌症、牛皮癣，以及伤口愈合受损。它也是 对于设计更好的培养条件以生产大型 用于移植和工程目的的正常人体细胞的数量。

DESCRIPTION: (Adapted from the applicant's abstract) - The long-term goal of the proposal is to understand the molecular regulatory mechanisms that maintain the cellular proliferative capacity and the signals that trigger the cells to cease proliferation. The principal investigator believes that understanding transcriptional regulation during the transition from proliferation to differentiation is an important step toward elucidation of these regulatory processes. The principal investigator has recently discovered a novel zinc finger protein, basonuclin. Basonuclin is present in the nuclei of keratinocytes in the proliferative compartment of epidermis and the germ cells in the seminiferous tubules of testis. The principal investigator hypothesizes that basonuclin is a transcription factor participating in the regulation of cellular proliferative potential (capability). Specific Aim 1 is to demonstrate that basonuclin is a DNA binding protein with sequence specificity. The approach is to select these targets from a pool of random sequences by electrophoresis mobility shift assay (EMSA) using zinc finger peptides. Specific Aim 2 will identify the genes whose promoters contain the basonuclin target sequence. The principal investigator aims to identify the genes that are controlled by basonuclin and thereby to understand the regulatory network in which basonuclin is a part. The principal investigator will search the DNA data base for promoters that contain the target sequence of basonuclin and to analyze the genomic fragments that are selected for their ability to bind to the bacterial produced full length basonuclin. Specific Aim 3 will map basonuclin's binding sites on mitotic chromosomes. The association of basonuclin with mitotic chromatin is highly unusual. The principal investigator aims to identify the mitotic chromosomes that contain basonuclin and to clone these binding sites, which may or may not be the same binding sites present on the bare DNA (lacking histones) that is studied in Aim 2. The virtue of molecules such as basonuclin in the study of biochemical and genetic regulation of cellular events is that it is a regulator that is also regulated. By studying it, the principal investigator expects to gain knowledge not only about basonuclin, but also about the regulatory hierarchy in which it is a part. The principal investigator believes that the proposed study of the biological function of basonuclin will enhance the knowledge about the molecular regulation of cellular proliferative potential. This knowledge is needed, he believes, to devise better medical treatment for proliferation-related abnormalities in the skin, such as cancer, psoriasis, and impaired wound healing. It is also essential for designing better culture conditions to produce large quantities of normal human cells for grafting and engineering purposes.