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首页> 外文期刊>Journal of Molecular Biology >Infectious human papillomavirus type 18 pseudovirions
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Infectious human papillomavirus type 18 pseudovirions

机译：感染性人乳头瘤病毒18型假病毒颗粒

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摘要

Human papillomavirus type 18 (HPV18) capsid proteins L1 and L2, synthesised in mammalian cells using recombinant vaccinia viral expression vectors, are transported to the nucleus and assembled into virus-like particles. When 293T cells, which express SV40 T antigen, were transfected with plasmid DNAs containing an SV40 origin of replication then infected with vaccinia viral vectors encoding L1 and L2, plasmid DNA was encapsidated into the particles. The DNAs ranged in size from 5.4 to 7.9 kb. By encapsidating plasmids containing either the beta-galactosidase gene or the puromycin-resistance gene, the pseudovirions were shown to be infectious in that they could transfer beta-galactosidase activity or confer resistance to puromycin to a number of cell types, indicating that the uptake and decapsidation of HPV particles are not the main determinants of cell type specificity of HPV. Episomal HPV16 DNA in a cervical keratinocyte line could also be encapsidated. Further investigation showed that DNA encapsidation is independent of HPV DNA sequences and of T antigen-mediated plasmid DNA replication. Instead, the minor capsid protein, L2, was found to be attached to plasmid mini-chromosomes extracted from these cells, suggesting a role for L2 in encapsidation. Consistent with this, the L1 protein alone was unable to encapsidate DNA, although it was able to form virus-like particles. The results suggest that intracellular episomal DNAs of suitable size can be encapsidated by the HPV18 L1 and L2 proteins without the need of any HPV packaging signal, and reintroduced into cells. (C) 1998 Academic Press. [References: 41]
机译：使用重组痘苗病毒表达载体在哺乳动物细胞中合成的人乳头瘤病毒18型（HPV18）衣壳蛋白L1和L2被转运到细胞核并组装成病毒样颗粒。当用含有SV40复制起点的质粒DNA转染表达SV40 T抗原的293T细胞，然后用编码L1和L2的牛痘病毒载体感染时，质粒DNA被衣壳化。 DNA的大小从5.4到7.9 kb不等。通过衣壳化含有β-半乳糖苷酶基因或嘌呤霉素抗性基因的质粒，假病毒颗粒被证明具有感染性，因为它们可以将β-半乳糖苷酶活性转移或赋予嘌呤霉素许多细胞类型抗性，表明其摄取和HPV颗粒的去壳化不是HPV细胞类型特异性的主要决定因素。宫颈角质形成细胞系中的游离型HPV16 DNA也可以被衣壳化。进一步的研究表明，DNA衣壳化与HPV DNA序列和T抗原介导的质粒DNA复制无关。取而代之的是，发现次要衣壳蛋白L2附着在从这些细胞中提取的质粒微型染色体上，表明L2在衣壳化过程中发挥了作用。与此相符的是，尽管L1蛋白能够形成病毒样颗粒，但它本身却不能包裹DNA。结果表明，HPV18 L1和L2蛋白可以包裹适当大小的细胞内游离DNA，而无需任何HPV包装信号，然后重新引入细胞。 （C）1998年学术出版社。 [参考：41]

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