Biological Function of Basonuclin

描述(由申请人提供)：一项关于生物功能的研究 巴索努克林。增加核糖体产量的能力对 细胞增殖。角质形成干细胞尤其如此。 因为它们必须持续分裂以维持正常的内稳态 组织，并且必须能够提高分裂的速度，以应对伤害。 Basonuclin是一种120 kDa的锌指蛋白，可能是一种转录蛋白 从基础水平增强核糖体RNA转录的调节因子。 Basonuclin与先前描述的rRNA转录因子不同 因为它的表达是细胞类型特异性和增殖相关的。我们的 假设碱性核蛋白的调节影响rRNA转录和 角质形成细胞中核糖体的产生，这种调节在 维持角质形成细胞的增殖能力。为了测试这一点 假设，我们提出了以下三个具体目标： 具体目标1，我们将通过系统突变和一种 共转染实验，将rRNA基因启动子中的序列(S) 这是Basonuclin作用所必需的。我们还将使用突变技术来定义 对Basonuclin作为一种蛋白质的功能至关重要的蛋白质结构域 RRNA转录激活剂。特异靶2，除核糖体RNA外 基因，Basonuclin可能调节另一组基因。我们假设这是 第二类包括编码核糖体蛋白的基因。我们将对此进行测试 通过确定basonuclin是否与启动子相互作用而提出假说 核糖体蛋白基因以及它是否影响其转录。具体目标3， 我们将构建表达显性-负性突变的转基因小鼠 基底角质形成细胞中的碱性核蛋白。我们会检查这个突变体是否能 抑制体内基底角质形成细胞Poll转录 这种抑制对正常发育和维持的影响 表皮。

DESCRIPTION (provided by the applicant): A study of the biological function of basonuclin. The ability to increase ribosome production is essential for cellular proliferation. This is particularly true for keratinocyte stem cells since they must divide continuously to maintain the normal homeostasis of the tissue and must be able to increase the rate of division in response to injury. Basonuclin, a zinc finger protein of 120 kDa, is likely to be a transcriptional regulator that enhances transcription of ribosomal RNA from a basal level. Basonuclin differs from the previously described rRNA transcription factors because its expression is cell type-specific and proliferation-related. Our hypothesis is that regulation of basonuclin affects rRNA transcription and ribosome production in keratinocytes and this regulation is important in maintaining the proliferative ability of keratinocytes. To test this hypothesis, we propose the following three specific aims: Specific Aim 1, We will investigate, by systematic mutagenesis and a co-transfection assay, the sequence (s) in the promoter of rRNA gene that is required for the action of basonuclin. We will also use mutagenesis to define the protein domains that are important for the function of basonuclin as an rRNA transcriptional activator. Specific Aim 2, In addition to ribosomal RNA genes, basonuclin may regulate another group of genes. We hypothesize that this second group includes genes encoding ribosomal proteins. We will test this hypothesis by determining if basonuclin interacts with the promoters of ribosomal protein genes and if it affects their transcription. Specific Aim 3, We will construct transgenic mice that express a dominant-negative mutation of basonuclin in the basal keratinocytes. We will examine if this mutant can inhibit Poll transcription in the basal keratinocytes in vivo and what are the effects of this inhibition on the normal development and maintenance of the epidermis.