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目的人乳头瘤病毒（Ｈｕｍａｎｐａｐｉｌｏｍａｖｉｒｕｓ，ＨＰＶ）１６型早期启动子Ｐ９７控制着病毒癌基因的表达。为了观测长控制区（Ｌｏｎｇｃｏｎｔｒｏｌｒｅｇｉｏｎ，ＬＣＲ）上ＹＹ１结合位点的破坏对病毒转化能力的影响。方法构建了带有自然发生突变ＬＣＲ序列的ＨＰＶ１６全基因质粒，利用ＥＭＳＡ和荧光素酶实验检测小鼠纤维细胞胞核内源性ＹＹ１蛋白存在和功能状态；将标准及突变的ＨＰＶ１６毒株转染至Ｃ１２７细胞进行软琼脂糖培养基生长实验。结果与上皮类细胞相似，在Ｃ１２７胞核提取物中可检出ＹＹ１蛋白，并且具有良好的ＤＮＡ结合功能和Ｐ９７活性抑制作用。转化实验显示ＹＹ１位点突变ＨＰＶ１６毒株可在软琼脂糖培养基上形成２～１０倍量多的生长集落。结论细胞转录调节因子ＹＹ１存在于啮齿类动物纤维细胞胞核中；ＬＣＲ上ＹＹ１结合位点的破坏可在完整基因组范围内明显增强病毒对小鼠纤维细胞的转化能力。
Objective Human papillomavirus (HPV) 16 type early promoter P97 controls the expression of viral oncogenes. In order to observe the effect of the disruption of the YY1 binding site on the long-term control region (LCR) on the virus transformation ability. Methods HPV16 genomic plasmid with naturally occurring LCR sequence was constructed. The presence and functional status of endogenous YY1 protein in the nuclei of mouse fibroblasts were detected by EMSA and luciferase assays. The standard and mutant HPV16 strains were transfected C127 cells were grown in soft agar medium. Results Similar to epithelial cells, YY1 protein was detected in C127 nuclear extracts and had good DNA binding and P97 activity inhibition. Transformation experiments showed that the YY1 mutant HPV16 strain can grow 2-10 times more colonies in soft agarose medium. Conclusions The transcriptional regulator YY1 is found in the nuclei of rodent fibroblasts. The disruption of the YY1 binding site on LCR can significantly enhance the ability of the virus to transfect mouse fibroblasts in the complete genome.