DIFFERENTIATION-INDUCED AND CONSTITUTIVE TRANSCRIPTION OF HUMAN PAPILLOMAVIRUS TYPE-31B IN CELL-LINES CONTAINING VIRAL EPISOMES
DIFFERENTIATION-INDUCED AND CONSTITUTIVE TRANSCRIPTION OF HUMAN PAPILLOMAVIRUS TYPE-31B IN CELL-LINES CONTAINING VIRAL EPISOMES
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DOI：
10.1128/jvi.66.10.6070-6080.1992
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发表时间：
1992-10-01
影响因子：
5.4
通讯作者：
LAIMINS, LA
中科院分区：
文献类型：
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作者：
HUMMEL, M;HUDSON, JB;LAIMINS, LA
人乳头瘤病毒 (HPV) 的生产生命周期中病毒基因的表达与上皮细胞的分化程序紧密相关。我们已经检查了 HPV 的转录作为体外器官筏培养系统中分化的功能，该系统允许上皮在空气-液体界面处分层。当含有 HPV 31b 型 (HPV31b) 游离型拷贝的 CIN612 细胞在筏培养物中分层时，它们以与宫颈上皮内瘤变 I 活检病变中所见的组织学相似的方式分化。在 CIN612 细胞的单层培养物中，鉴定出两个主要的 1.7 kb 多顺反子 HPV31b 转录物，它们编码 (i) E6、E7、E1^E4 和 E5 以及 (ii) E6*、E7、E1 E4 和 E5。这些 RNA 起始于病毒上游调控区的启动子 P97。在筏培养物中分化后，P97 起始的 RNA 的相对丰度没有变化。相反，编码E1、E4融合蛋白和E5的1.3-kb RNA的表达被发现在分化后显着增加。该转录本在 E7 基因 (P742) 内的一个新启动子处启动。因此，这些研究鉴定了在整个复层上皮中具有活性的组成型病毒启动子以及在上皮细胞分化时诱导的新型启动子。
The expression of viral genes during the productive life cycle of human papillomaviruses (HPV) is tightly coupled to the differentiation program of epithelial cells. We have examined transcription of HPV as a function of differentiation in an in vitro organotypic raft culture system which allows for epithelial stratification at the air-liquid interface. When CIN612 cells, which contain episomal copies of HPV type 31b (HPV31b), were allowed to stratify in raft cultures, they differentiated in a manner which was histologically similar to that seen in a cervical intraepithelial neoplasia I biopsy lesion. In monolayer cultures of CIN612 cells, two major polycistronic HPV31b transcripts of 1.7 kb which encode (i) E6, E7, E1^E4, and E5 and (ii) E6*, E7, E1 E4, and E5 were identified. These RNAs initiated at a promoter, P97, in the upstream regulatory region of the virus. Following differentiation in raft cultures, the relative abundance of RNAs initiated at P97 was unchanged. In contrast, the expression of a 1.3-kb RNA encoding an E1 E4 fusion protein and E5 was found to increase substantially following differentiation. This transcript was initiated at a novel promoter within the E7 gene (P742). These studies have therefore identified a constitutive viral promoter which is active throughout stratified epithelium as well as a novel promoter which is induced upon epithelial cell differentiation.