IMMUNOLOGICAL CHARACTERIZATION OF CELLS

乳头瘤病毒(PV)是一种令人着迷的病原体，它能引起许多 常见的病变包括皮肤、肛门和宫颈 有疣。人类多发性硬化症与阴茎、外阴和 宫颈癌。 乳头瘤病毒具有相对较小的(8000)碱基对DNA 编码10-12个蛋白质的基因组，并将其作为范例 基因调控、DNA复制和致癌 变身。牛(B)光伏是应用最广泛的遗传基因 用于检查光伏功能的模型。一个千碱基的区域 BPV包含调控TIS表达的DNA序列 基因组。与几个启动子一起，该区域包括一个顺式- 作用增强子，刺激转录不依赖于 方位或位置。该增强剂的激活依赖于 在BPVE2阅读框的一个基因产物上。E2一直是 被证明是一种序列特异性DNA结合蛋白，识别 这是所有光伏基因组共有的基序。此处介绍的数据 证明增强子活性是由E2 DNA提供的 与异源启动子和E2基因连锁的结合位点 产品。 拟议的实验包括表征 E2的结构、形式、翻译后状态和稳定性 BPV感染细胞中多克隆和单克隆的蛋白质 将使用抗体。雌二醇与其DNA识别的结合 将对场地进行研究。2的职能组织将是 由突变分析确定，并将与其相关 结构、生化特性和反式激活能力。 它与细胞因子相互作用的假说 将检测转录增强。这些研究将 阐明对PV基因调控的进一步理解以及 提供对基因控制的增强子机制的见解。

Papillomaviruses (PVs) are fascinating pathogens that cause many common lesions including cutaneous, anogenital, and cervical warts. Human PVs are closely associated with penile, vulvar, and cervical cancers. Papillomaviruses have relatively small (8000) base pair DNA genomes that encode 10-12 proteins and lends itself as a paradigm of gene regulation, DNA replication, and oncogenic ransformation. The bovine (B) PV is the most widely used genetic model for examination of PV functions. A one kilobase region of BPV contains DNA sequences that regulate expression of tis genome. Along with several promoters, this region includes a cis- acting enhancer that stimulates transcription independent of orientation or position. Activation of this enhancer is dependent on a gene product of the BPV E2 reading frame. E2 has been shown to be a sequence-specific DNA binding protein, recognizing a motif common to all PV genomes. Data described here demonstrate that enhancer activity is provided by the E2 DNA binding site linked to a heterologous promoter and the E2 gene product. The proposed experiments involve characterization of the structure, form, post-translational state, and stability of the E2 protein in BPV-infected cells for which polyclonal and monoclonal antibodies will be used. The binding of E2 to its DNA recognition site will be studied. The funcitonal organization of E2 will be determined by mutational analysis and will be correlated with its structure, biochemical properties, and transactivating capacity. The hypothesis that it interacts with cellular factors for transcriptional enhancement will be examined. These studies will elucidate further understanding of PV gene regulation as well as provide insights into the enhancer mechanism of gene control.