Molecular Inflammation Research Group, Department of Medical Biology, Faculty of Health Sciences, University of Tromsø, The Arctic University of Norway, Tromsø, Norway.
Present address: Tumor Biology Research Group, Department of Medical Biology, Faculty of Health Sciences, University of Tromsø, The Arctic University of Norway, Tromsø, Norway.
Virol J. 2020 Apr 19;17(1):54. doi: 10.1186/s12985-020-01317-x.
Merkel cell polyomavirus (MCPyV) is a human polyomavirus that establishes a life-long harmless infection in most individuals, with dermal fibroblasts believed to be the natural host cell. However, this virus is the major cause of Merkel cell carcinoma (MCC), an aggressive skin cancer. Several MCPyV variants with polymorphism in their promoter region have been isolated, but it is not known whether these differences affect the biological properties of the virus.
Using transient transfection studies in human dermal fibroblasts and the MCC cell line MCC13, we compared the transcription activity of the early and late promoters of the most commonly described non-coding control region MCPyV variant and six other isolates containing specific mutation patterns.
Both the early and late promoters were significantly stronger in human dermal fibroblasts compared with MCC13 cells, and a different promoter strength between the MCPyV variants was observed. The expression of full-length large T-antigen, a viral protein that regulates early and late promoter activity, inhibited early and late promoter activities in both cell lines. Nonetheless, a truncated large T-antigen, which is expressed in virus-positive MCCs, stimulated the activity of its cognate promoter.
The promoter activities of all MCPyV variants tested was stronger in human dermal fibroblasts, a cell line that supports viral replication, than in MCC13 cells, which are not permissive for MCPyV. Truncated large T-antigen, but not full-length large T-antigen stimulated viral promoter activity. Whether, the difference in promoter strength and regulation by large T-antigen may affect the replication and tumorigenic properties of the virus remains to be determined.
默克尔细胞多瘤病毒(MCPyV)是一种人类多瘤病毒,在大多数个体中建立长期无害的感染,真皮成纤维细胞被认为是其自然宿主细胞。然而,这种病毒是 Merkel 细胞癌(MCC)的主要原因,MCC 是一种侵袭性皮肤癌。已经分离出几种在其启动子区域具有多态性的 MCPyV 变体,但尚不清楚这些差异是否影响病毒的生物学特性。
我们使用瞬时转染研究在人真皮成纤维细胞和 MCC 细胞系 MCC13 中,比较了最常描述的非编码控制区 MCPyV 变体和其他六个包含特定突变模式的分离物的早期和晚期启动子的转录活性。
与 MCC13 细胞相比,早期和晚期启动子在人真皮成纤维细胞中均显著增强,并且观察到 MCPyV 变体之间的不同启动子强度。全长大 T 抗原的表达,一种调节早期和晚期启动子活性的病毒蛋白,抑制了两种细胞系中的早期和晚期启动子活性。尽管如此,在病毒阳性 MCC 中表达的截短大 T 抗原刺激了其同源启动子的活性。
与不允许 MCPyV 复制的 MCC13 细胞相比,在支持病毒复制的人真皮成纤维细胞中,测试的所有 MCPyV 变体的启动子活性均更强。截短的大 T 抗原而非全长大 T 抗原刺激病毒启动子活性。启动子强度的差异和大 T 抗原的调节是否会影响病毒的复制和致瘤特性仍有待确定。