单纯疱疹病毒DNA在受感染人类细胞中的物理状态。

The physical state of herpes simplex virus DNA in infected human cells.

作者信息

Hirai K

出版信息

Microbiol Immunol. 1979;23(8):749-61. doi: 10.1111/j.1348-0421.1979.tb00517.x.

DOI:10.1111/j.1348-0421.1979.tb00517.x

Abstract

Treatment of herpes simplex virus type 1 (HSV-1)-infected human embryo lung (HEL) cells with phosphonoacetic acid (PAA) resulted in complete inhibition of HSV DNA replication. DNA was extracted from PAA-treated HEL cells infected with HSV-1 and centrifuged in a neutral CsCl density gradient. The HSV DNA sequences in the nuclei of PAA treated cells at 24 hr post infection banded at the same density as free HSV DNA (1.725 g/cm3), but a significant amount of viral DNA sequences were detected in the regions of cell DNA (1.700 g/cm3) as well as in the intermediate fractions as determined by hybridization with 3H HSV complementary RNA. The viral DNA sequences of lower deisntiy did not change in density by recentrifugation in a CsCl density gradient, but did change to the density of free viral DNA after treatment with EcoR1 restriction endonuclease. When the DNA from the nuclei of PAA treated cells was analyzed in an alkaline glycerol gradient, more than 95% of the viral DNA sequences were found in the free viral DNA fractions. Since the viral and cellular hybrid DNA represented approximately 33% of the total viral DNA sequences, it is concluded that some of the HSV DNA sequences in PAA treated, infected cells are associated with cell DNA by alkali-labile bonds.

摘要

用膦甲酸（PAA）处理单纯疱疹病毒1型（HSV-1）感染的人胚肺（HEL）细胞，可完全抑制HSV DNA复制。从感染HSV-1的经PAA处理的HEL细胞中提取DNA，并在中性CsCl密度梯度中离心。感染后24小时，经PAA处理的细胞细胞核中的HSV DNA序列在与游离HSV DNA相同的密度（1.725 g/cm³）处形成条带，但通过与³H HSV互补RNA杂交确定，在细胞DNA区域（1.700 g/cm³）以及中间组分中也检测到大量病毒DNA序列。密度较低的病毒DNA序列在CsCl密度梯度中再次离心后密度不变，但在用EcoR1限制性内切酶处理后变为游离病毒DNA的密度。当在碱性甘油梯度中分析经PAA处理的细胞细胞核中的DNA时，超过95%的病毒DNA序列存在于游离病毒DNA组分中。由于病毒与细胞的杂交DNA约占病毒DNA序列总数的33%，因此得出结论，在经PAA处理的感染细胞中，一些HSV DNA序列通过碱不稳定键与细胞DNA相关联。