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HPV-2是引起皮肤寻常疣的常见HPV型别,病毒E2蛋白可抑制病毒早期启动子的活性。我们曾经报道来自一例巨大寻常疣患者的HPV-2突变E2蛋白对病毒早期启动子活性的抑制作用明显减弱,该E2蛋白在其C末端的DNA结合区域带有A338V的点突变。本研究利用原核表达系统表达纯化了突变E2(A338V)和HPV-2原毒株的羧基端和全长蛋白。电泳迁移率实验结果显示,E2蛋白可与带有E2蛋白特异性结合位点的寡核苷酸探针形成复合物,突变E2蛋白比原毒株E2蛋白的DNA结合能力强。这提示DNA结合能力的增强可能为E2蛋白对病毒启动子活性影响的分子基础,与患者出现罕见巨大寻常疣这一临床表型关联。
HPV-2 is a common HPV type that causes verruca vulgaris. The virus E2 protein inhibits the early promoter activity of the virus. We have reported that the inhibition of the early promoter activity of the virus by the HPV-2 mutant E2 protein from a patient with giant verruca vulgaris significantly diminishes the point-mutation of the A338V in its C-terminal DNA-binding region. In this study, the prokaryotic expression system was used to purify the carboxyl-terminal and full-length proteins of mutant E2 (A338V) and HPV-2 virulent strains. The electrophoretic mobility shift assay showed that the E2 protein could form a complex with the oligonucleotide probe with the specific binding site of E2 protein. The mutant E2 protein has stronger DNA binding ability than the E2 protein of the original strain. This suggests that increased DNA binding may be the molecular basis of the effect of E2 protein on viral promoter activity in association with the clinical phenotype of patients with unusually large verruca vulgaris.