Characterization of late gene transcripts expressed during vegetative replication of human papillomavirus type 31b
Characterization of late gene transcripts expressed during vegetative replication of human papillomavirus type 31b
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DOI：
10.1128/jvi.71.7.5161-5172.1997
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发表时间：
1997-07-01
影响因子：
5.4
通讯作者：
Meyers, C
中科院分区：
文献类型：
--
作者：
Ozbun, MA;Meyers, C
人乳头瘤病毒 (HPV) 是肛门生殖器癌症的病原体，缺乏有效的体外系统来研究分化依赖性病毒生命周期，这阻碍了大多数病毒转录和基因表达的研究，CIN-612 克隆 9E 细胞系潜在地维持 HPV 31b 型 (NPV31b) 的附加型拷贝。 HPV31b 的完整复制生命周期可以通过使用器官型研究（筏）养殖系统。使用raft系统的研究中已经描述了许多剪接的HPV31b早期基因转录本和两个晚期基因转录本，使用该系统已经对HPV31b早期启动子P-97和分化诱导启动子P-742进行了表征。在本研究中，我们使用raft系统分析了HPV31b晚期基因转录本在病毒生命周期中的时间表达模式，晚期RNA的表达在提升到最高水平后的第12天达到峰值。气液界面；然后水平在第 16 天急剧下降。晚期 RNA 表达的峰值与筏组织中病毒颗粒的出现一致。我们鉴定了具有编码晚期基因产物潜力的转录物，包括 19 个含有 L1 区域的 RNA 和 4 个含有 E5b 和 L2 开放阅读框的 RNA。我们还发现了两个新启动子的证据，含有 L1 和 L2 的 RNA 的转录起始于早期启动子上游的区域。此外，晚期基因 RNA 也被通过使用 E4 阅读框中的启动子进行转录。
Human papillomaviruses (HPVs) are etiologic agents of anogenital cancers, The lack of an efficient in vitro system with which to study the differentiation-dependent viral life cycle has impeded most investigations of viral transcription and gene expression, The CIN-612 clone 9E cell line latently maintains episomal copies of HPV type 31b (NPV31b), The complete replicative life cycle of HPV31b, can be studied by using the organotypic (raft) culture system. A number of spliced HPV31b early gene transcripts and two late gene transcripts have been described in studies using the raft system, An HPV31b early promoter, P-97, and a differentiation-induced promoter, P-742, have been characterized by using this system, In this study, we used the raft system to analyze the temporal expression patterns of HPV31b late gene transcripts during the viral life cycle, The expression of late RNAs peaked at day 12 after lifting to the air-liquid interface; the levels then declined dramatically by day 16. The peak of late RNA expression was coincident with the appearance of virus particles in the raft tissues, We characterized transcripts with the potential to encode late gene products, including 19 RNAs containing the L1 region and 4 RNAs containing the E5b and L2 open reading frames, We also found evidence for two novel promoters, Transcription of both L1- and L2-containing RNAs initiated at a region upstream of the early promoter, In addition, late gene RNAs were also transcribed by using a promoter in the E4 reading frame.