HPV 6 TRANSCRIPTION

描述(改编自申请人的摘要)：人 乳头瘤病毒(HPV)是一种小的DNA病毒，可引起良性病变 鳞状上皮。目前已知的HPV基因类型有60多种，其中 大多数生殖器感染含有6、11、16和18型DNA。 其中11例最常与良性病变有关。然而，这两个人 类型也与Buschke-Lowenstein疣密切相关 癌，一种侵袭性生长的非转移性病变。高度的 HPV6诱导的外生性良性肿瘤，尖锐湿疣， 疣状癌的侵袭性表型提示 病毒在感染上皮细胞异常增殖中的重要作用 细胞。最近的研究表明，人乳头瘤病毒与人乳头瘤病毒 6基因表达与上皮细胞终末分化。虽然 高危HPV类型的基因表达机制是众所周知的， 人们对类似的HPV6基因是如何调控的知之甚少。在……里面 事实上，似乎E6和E7癌基因的表达可能受到调控 不同的生殖器HPV类型具有不同的致癌潜能。在……里面 与HPV 16和18，即所谓的高致癌类型相比，假定 人乳头瘤病毒6型的基因调控序列似乎超出了非编码 区域(URR)插入E6、E7和E1开放阅读框，因为 转录启动子已经被映射到下游的3个不同的位置 E6ORF的5‘端。此应用程序的初始目标是 了解不同HPV6启动子之间的相互作用，他们的 病毒和细胞蛋白的调节，以及它们在 人类角质形成细胞永生化。 由于人乳头瘤病毒对上皮细胞有严格的趋向性， 在以下背景下研究HPV6启动子活性是至关重要的 自然的宿主细胞。携带HPV6调节片段的载体将 在瞬时转染法中用作转录模板。 上皮细胞种类繁多。由个别启动者产生的文本 将通过S1作图和引物延伸进行鉴定 人乳头瘤病毒6型核糖核酸分析不同语种间转录的比较研究 将进行细胞类型鉴定，以确定细胞特异性启动子(S)。 这些检测方法也将用于重组、缺失和 接头扫描HPV6调控区突变体以鉴定 参与病毒转录的反应元件。因为 乳头瘤病毒还编码几种调节蛋白，其作用是 不同的病毒蛋白对HPV6转录的影响将被分析。角色 HPV6调节区对永生化潜能的影响将被研究 通过构建HPV6和18之间的重组体并检测它们的能力 使人类包皮角质形成细胞永生化。通过这些研究， 申请人希望阐明HPV6基因调控的机制。

DESCRIPTION (Adapted from the applicant's abstract): Human papillomaviruses (HPV) are small DNA viruses which induce benign lesions of squamous epithelia. More than 60 HPV genotypes are known, and the vast majority of genital infections contain DNA types 6, 11, 16 and 18. HPV 6 and 11 are most often associated with benign lesions. However, these two types are also closely associated with Buschke-Lowenstein verrucous carcinoma, an invasively growing, non-metastasizing lesion. The highly exophytic nature of HPV 6 induced benign tumors, the condyloma acuminata, and the aggressive phenotype of the verrucous carcinomas suggest an important viral role in the aberrant proliferation of infected epithelial cells. Recent studies have suggested an intimate relationship between HPV 6 gene expression and epithelial cell terminal differentiation. Although the mechanism of gene expression in high-risk HPV types is well understood, little is known about how the analogous HPV 6 genes are regulated. In fact, it appears that expression of E6 and E7 oncogenes may be regulated differently in genital HPV types of different oncogenic potential. In contrast to HPV 16 and 18, the so-called high oncogenic types, the putative gene regulatory sequences of HPV 6 appear to extend beyond the non-coding region (URR) into the E6, E7 and E1 open reading frames, because transcriptional promoters have been mapped to 3 distinct sites downstream of the 5' end of E6 ORF. The initial goal of this application is to understand the interaction among different HPV 6 promoters, their regulation by viral and cellular proteins, and their role in the immortalization of human keratinocytes. Because human papillomaviruses have a strict tropism for epithelial cells, it is essential that HPV 6 promoter activities be studied in the context of the natural host cell. Vectors carrying the HPV 6 regulatory fragment will be used as transcriptional templates in transient transfection assays in a variety of epithelial cells. Transcripts arising from individual promoters in transfected cells will be identified by S1 mapping and primer extension analysis of HPV 6 RNA. Comparative studies of transcription in different cell types will be carried out to identify cell-specific promoter(s). These assays will also be used in analysis of recombinant, deletion and linker-scanning mutants of the HPV 6 regulatory region to identify responsive elements involved in viral transcription. Because papillomaviruses also encode several regulatory proteins, the effect of different viral proteins on HPV 6 transcription will be analyzed. The role of HPV6 regulatory region on the immortalization potential will be studied by constructing recombinants between HPV6 and 18 and testing their ability to immortalize human foreskin keratinocytes. Through these studies, the applicant hopes to elucidate the mechanism of HPV 6 gene regulation.