Nucleosome Positioning on Episomal Human Papillomavirus DNA in Cultured Cells.
Nucleosome Positioning on Episomal Human Papillomavirus DNA in Cultured Cells.
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DOI：
10.3390/pathogens10060772
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发表时间：
2021-06-19
期刊：
影响因子：
--
通讯作者：
Aoki D
中科院分区：
文献类型：
--
作者：
Murakami I;Iwata T;Morisada T;Tanaka K;Aoki D
几种人乳头瘤病毒 (HPV) 与宫颈癌的发生有关。当受感染的宿主角质形成细胞从上皮基底层迁移到棘层时，HPV DNA 合成会增加，但分子机制尚不清楚。核小体定位通过允许转录因子接近启动子来启动转录来影响各种细胞过程，例如 DNA 复制和修复。在这项研究中，在稳定转染 HPV16 或 HPV18 基因组的正常永生化角质形成细胞 (NIKS) 中研究了病毒染色质上的核小体定位，以确定是否与病毒生命周期存在关联。使用针对 HPV16 和 HPV18 的探针通过 Southern 印迹分析微球菌核酸酶处理的 DNA，并使用实时 PCR 进行核小体扫描分析进行定量，结果显示 140-200 个碱基对的单核小体大小的片段，这些片段在病毒基因组中的位置根据细胞是否正在增殖或分化而变化。值得注意的是，增殖和分化宿主细胞中核苷酸 110 周围区域的变化对于 HPV16 和 HPV18 来说是常见的。我们的研究结果表明，宿主细胞分化过程中病毒DNA上核小体位置的变化是病毒生命周期中的重要调控事件。
Several human papillomaviruses (HPV) are associated with the development of cervical carcinoma. HPV DNA synthesis is increased during the differentiation of infected host keratinocytes as they migrate from the basal layer of the epithelium to the spinous layer, but the molecular mechanism is unclear. Nucleosome positioning affects various cellular processes such as DNA replication and repair by permitting the access of transcription factors to promoters to initiate transcription. In this study, nucleosome positioning on virus chromatin was investigated in normal immortalized keratinocytes (NIKS) stably transfected with HPV16 or HPV18 genomes to determine if there is an association with the viral life cycle. Micrococcal nuclease-treated DNA analyzed by Southern blotting using probes against HPV16 and HPV18 and quantified by nucleosome scanning analysis using real-time PCR revealed mononucleosomal-sized fragments of 140–200 base pairs that varied in their location within the viral genome according to whether the cells were undergoing proliferation or differentiation. Notably, changes in the regions around nucleotide 110 in proliferating and differentiating host cells were common to HPV16 and HPV18. Our findings suggest that changes in nucleosome positions on viral DNA during host cell differentiation is an important regulatory event in the viral life cycle.
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