HPV 6 TRANSCRIPTION

描述（改编自申请人摘要）：人 乳头状瘤病毒（HPV）是小DNA病毒，其诱导良性病变， 鳞状上皮 已知有60多种HPV基因型， 大多数生殖器感染含有DNA类型6，11，16和18。 HPV 6 11例最常与良性病变相关。 但这两 类型也与Buschke-Lowenstein疣状 癌，一种侵袭性生长的非转移性病变。 高度 HPV 6的外生性诱导良性肿瘤，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，HPV与 6基因表达与上皮细胞终末分化。 虽然 高风险HPV类型中基因表达的机制已被充分理解， 关于类似的HPV 6基因是如何被调节的知之甚少。 在 事实上，E6和E7癌基因的表达似乎可以被调节， 不同的生殖器HPV类型具有不同的致癌潜力。 在 与所谓的高致癌类型HPV 16和18相比， HPV 6的基因调控序列似乎延伸到非编码区以外， E6、E7和E1开放阅读帧，因为 转录启动子已定位于下游3个不同位点 E6 ORF的5'端。 本应用程序的初始目标是 了解不同HPV 6启动子之间的相互作用， 病毒和细胞蛋白的调节，以及它们在 人角质形成细胞的永生化。 因为人乳头瘤病毒对上皮细胞有严格的嗜性， 在以下背景下研究HPV 6启动子活性是至关重要的： 自然宿主细胞。 携带HPV 6调节片段的载体将 在瞬时转染测定中用作转录模板， 各种上皮细胞。 由单个启动子产生的转录本 将通过S1作图和引物延伸鉴定转染细胞中的 HPV 6 RNA分析。 不同语言转录的比较研究 将进行细胞类型鉴定以鉴定细胞特异性启动子。 这些测定也将用于分析重组、缺失和 HPV 6调控区的接头扫描突变体，以鉴定 参与病毒转录的应答元件。 因为 乳头瘤病毒还编码几种调节蛋白， 将分析不同病毒蛋白对HPV 6转录的影响。的作用 研究HPV 6调控区对永生化潜能的影响 通过构建HPV 6和HPV 18之间的重组体并检测其能力， 使人类包皮角质细胞永生化。 通过这些研究， 申请人希望阐明HPV 6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.