ROLE OF PAPILLOMAVIRUS DNA IN CELL TRANSFORMATION

虽然人乳头瘤病毒与恶性肿瘤有关， 疾病，对这些病毒的研究一直受到阻碍，因为缺乏 合适的组织培养系统。 为了理解 乳头瘤病毒基因组在细胞转化，实验已经 设计用于将功能分配给特定的开放阅读框架， 在培养物中有活性的两种动物病毒的基因组的转化区; 牛乳头瘤病毒1型（BPV-1）和鹿乳头瘤病毒（DPV）。 研究将涉及（i）干扰开放阅读框架内的 转化区域并通过细胞转化测定效果，和（ii） 通过细菌表达载体合成编码的推定多肽 以确定它们是否在肿瘤中表达 细胞 BPV-1和DPV差异转化小鼠细胞。 BPV-1变换C127 而DPV不能转化C127细胞 细胞 已经有人提出实验来创建嵌合基因组， 这两种病毒通过体外和体内重组，并确定 三种细胞系的转化能力。 这些研究可以提供 乳头瘤病毒控制机制的信息，因为嵌合体将 代表了从这两个衍生的复制和转化域 病毒。 三个新的BPV分离物从从DNA获得的分子克隆， 将进一步表征形态学上不同的乳头乳头状瘤。 将测定这些病毒基因组在细胞培养物中的转化， 他们的基因组通过DNA测序进行分析。 丁酸钠抑制BPV-1转化小鼠细胞的生长， 降低BPV-1基因组拷贝数。 从处理的获得的克隆 培养物具有平坦的表型。 为了进一步研究这一点，获得了BPV-1 DNA 将对扁平回复突变体进行表征，以确定它们是否 也有人提出研究来确定衰变的动力学 以及肿瘤启动子是否可以重建 在扁平回复突变体中转化表型。 这些研究应该会产生 深入了解乳头瘤病毒的转化机制。

Although the human papillomaviruses have been implicated in malignant disease, research on these viruses have been hampered because of the lack of a suitable tissue culture system. To understand the role of the papillomavirus genome in cell transformation, experiments have been designed to assign function to specific open reading frames in the transforming region of the genomes of two animal viruses active in culture; bovine papillomavirus type 1 (BPV-1) and deer papillomavirus (DPV). Studies will involve (i) perturbation of open reading frames within the transforming region and assaying the effect by cell transformation and (ii) synthesis by bacterial expression vectors of putative polypeptides encoded within the transforming region to determine if they are expressed in tumor cells. BPV-1 and DPV differentially transform mouse cells. BPV-1 transforms C127 and NIH/3T3 but not Balb/3T3 cells whereas DPV is unable to transform C127 cells. Experiments have been proposed to create chimeric genomes between these two viruses by recombination in vitro and in vivo and determine the transforming ability on the three cell lines. These studies may provide information into papillomavirus control mechanisms since chimeras will represent replicating and transforming domains derived from these two viruses. Three new BPV isolates molecularly cloned from DNA obtained from morphologically distinct teat papillomas will be further characterized. These virus genomes will be assayed for transformation in cell culture and their genomes analyzed by DNA sequencing. Sodium butyrate inhibits the growth of BPV-1-transformed mouse cells and decreases the BPV-1 genome copy number. Clones obtained from treated cultures have a flat phenotype. To study this further, BPV-1 DNAs obtained from flat revertants will be characterized to determine if they have been mutated and studies have also been proposed to determine kinetics of decay of the viral sequences and whether tumor promoters can reestablish the transformed phenotype in flat revertants. These studies should yield insights into the mechanism of transformation by papillomaviruses.