ROLE OF PAPILLOMAVIRUS DNA IN CELL TRANSFORMATION

尽管人类乳头瘤病毒被认为与恶性肿瘤有关 疾病，对这些病毒的研究由于缺乏 合适的组织培养体系。要了解 乳头瘤病毒基因组在细胞转化中的实验研究 设计为将功能分配给 培养中活跃的两种动物病毒基因组的转化区； 牛乳头瘤病毒1型(BPV-1)和鹿乳头瘤病毒(DPV)。 研究将涉及(I)开放阅读框架的扰动 转化区域及细胞转化效果的测定和(Ii) 利用细菌表达载体合成可能编码的多肽 以确定它们是否在肿瘤中表达 细胞。 BPV-1和DPV可以不同地转化小鼠细胞。BPV-1改造C127 和NIH/3T3，而不是Balb/3T3细胞，而DPV不能转化C127 细胞。已经有人提出了创造嵌合基因组的实验 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。这些研究可能会提供 信息进入乳头瘤病毒控制机制，因为嵌合体将 表示从这两个域派生的复制和转换域 病毒。 从牛细小病毒DNA中分子克隆出三株新的牛细小病毒 在形态上不同的乳头状瘤将被进一步描述。 这些病毒基因组将在细胞培养和转化过程中进行检测 通过DNA测序分析它们的基因组。 丁酸钠抑制BPV-1转化的小鼠细胞的生长和 减少BPV-1基因组拷贝数。从处理后获得的克隆 培养物具有扁平的表型。为了进一步研究这一点，获得了BPV-1 DNA 将对扁平反转体进行表征，以确定它们是否已经 突变，也有人提出了确定衰变动力学的研究 以及肿瘤促进剂是否能重建 扁平反转突变体的转化表型。这些研究应该会产生 对乳头瘤病毒转化机制的洞察。

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.