Transcription activities of human papillomavirus type 11 E6 promoter-proximal elements in raft and submerged cultures of foreskin keratinocytes.
Transcription activities of human papillomavirus type 11 E6 promoter-proximal elements in raft and submerged cultures of foreskin keratinocytes.
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人乳头瘤病毒 11 型 E6 启动子近端元件在包皮角质形成细胞的筏和深层培养物中的转录活性。
DOI：
10.1128/jvi.71.11.8832-8840.1997
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发表时间：
1997
影响因子：
5.4
通讯作者：
Broker,TR
中科院分区：
文献类型：
--
作者：
Zhao,W;Chow,LT;Broker,TR
人乳头瘤病毒（HPV）复制，只有在分化的鳞状上皮疣和上皮筏培养生长在介质-空气界面。病毒编码的转录因子和宿主转录因子被认为负责抑制位于未分化基底细胞和副基底细胞上游调控区（URR）内的病毒增强子和启动子，同时上调它们在分化的棘细胞中的活性。使用重组逆转录病毒，我们急性转导新生儿包皮角质形成细胞（PHK）与lacZ报告基因驱动的野生型URR的低风险的HPV 11型或URR与个别突变的7个启动子近端元件，其中一些还没有以前的特点。β-半乳糖苷酶的活动中检测到淹没，增殖PHKs，也在分化的棘细胞，但不是在稳定状态下增殖的基底细胞，分层筏文化。特别是，突变的Oct 1，Sp1，或以前未知的启动子近端AP 1位点严重降低了报告活性，而突变的两个NF 1位点侧翼的Oct 1位点没有影响。这些结果表明在不同的培养条件下细胞转录因子谱的变化，并开始表征自然分化依赖性激活的URR。它们为疣中HPV表达模式提供了一种分子解释，并有助于验证上皮筏培养作为HPV调控元件遗传解剖的重要实验系统。
Human papillomaviruses (HPVs) replicate only in differentiated squamous epithelia in warts and in epithelial raft cultures grown at the medium-air interface. Virus-encoded and host transcription factors are thought to be responsible for repressing the viral enhancer and promoter located within the upstream regulatory region (URR) in the undifferentiated basal and parabasal cells while up-regulating their activities in the differentiated spinous cells. Using recombinant retroviruses, we acutely transduced neonatal foreskin keratinocytes (PHKs) with a lacZ reporter gene driven by the wild-type URR of the low-risk HPV type 11 or by a URR with individual mutations in seven promoter-proximal elements, some of which have not been characterized previously. Beta-galactosidase activities were detected in the submerged, proliferating PHKs and also in the differentiated spinous cells, but not in the steady-state proliferating basal cells, of stratified raft cultures. In particular, mutation of an Oct1, an Sp1, or a previously unknown promoter-proximal AP1 site severely reduced the reporter activity, whereas mutation of either of two NF1 sites flanking the Oct1 site had no effect. These results demonstrate changes in cellular transcription factor profiles under different culture conditions and begin to characterize the naturally differentiation-dependent activation of the URR. They provide one molecular explanation for the patterns of HPV expression in warts and help validate epithelial raft cultures as an important experimental system for genetic dissection of HPV regulatory elements.