IMMUNOLOGICAL CHARACTERIZATION OF CELLS

乳头瘤病毒（PV）是一种迷人的病原体， 常见病变包括皮肤、肛门生殖器和宫颈 疣。 人类PV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种蛋白质的基因组， 基因调控、DNA复制和致癌 转化。 牛（B）PV是最广泛使用的遗传疫苗。 检查PV功能的模型。 一个单核苷酸酶区域， BPV含有调节TIS表达的DNA序列， 基因组 沿着几个启动子，该区域包括顺式- 作用增强子，其刺激转录独立于 方向或位置。 这种增强子的激活依赖于 在BPV E2阅读框的基因产物上。 E2已经 显示为序列特异性DNA结合蛋白，识别 所有PV基因组共有的基序。 此处描述的数据 证明增强子活性由E2 DNA提供 与异源启动子和E2基因连接的结合位点 产品 拟议的实验涉及表征的 E2的结构、形式、翻译后状态和稳定性 BPV感染细胞中的蛋白质， 将使用抗体。 E2与其DNA识别的结合 网站将进行研究。 E2的职能组织将是 通过突变分析确定，并将与其 结构、生化特性和反式激活能力。 假设它与细胞因子相互作用， 将检查转录增强。 这些研究将 阐明进一步了解PV基因调控以及 提供了深入了解基因控制的增强子机制。

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.