Spector D, Pizer L I
J Virol. 1978 Jul;27(1):1-12. doi: 10.1128/JVI.27.1.1-12.1978.
The effect of herpes simplex virus (HSV) infection of mRNA metabolism was examined in a system where the fate of specific RNA sequence can be assayed. Adenovirus type 5-transformed rat embryo cell line 107 synthesizes adenovirus-specific RNA (ad-RNA), which functions in the cytoplasm as mRNA. We have utilized ad-RNA as a model for mRNA metabolism, and in a preliminiary study we characterized ad-RNA in the nucleus and cytoplasm by hybridization to filter-bound adenovirus DNA. The results indicated the as-RNA accumulates in the nucleus and that cytoplasmic polyadenylic acid [poly(A)]-containing ad-RNA turns over with a half-life of a few hours. Pulse-chase experiments confirmed these observations and a half-life of about h was determined for the poly(A)-containing cytoplasmic ad-RNA. A second class of ad-RNA remains in the nucleus, where it turns over with a longer hlaf-life (about 24 h). The infection of 107 cells by HSV was restricted at 37 degree C, giving a burst size of 5 PFU per cell and allowing continued host DNA synthesis. Protein synthesis was inhibited greater than 50% by 7 h after infection, and total RNA synthesis was 50% inhibited by 4 h after infection. During the first 8 h after infection, HSV has little effect on the rate of synthesis of ad-RNA as determined by hybridization of nuclear RNA samples, but,during the same period, HSV inhibits the accumulation of poly(A)-containing ad-RNA in the cytoplasm. The degree of this inhibition increases steadily throughout this period and reaches 60% by 6.5 to 8 h after infection. Nosignificant effect was seen on the accumulation of total cellular poly(A)-containing RNA. It was concluded from these experiments that HSV infection alters the metabolism of ad-RNA so as to prevent the normal appearance of the poly(A)-containing mRNA in the cytoplasm. The result for ad-RNA may not represent the behavior of total cellular poly(A)-containing RNA under conditions where infection is restricted.
在一个能够检测特定RNA序列命运的系统中，研究了单纯疱疹病毒（HSV）感染对mRNA代谢的影响。5型腺病毒转化的大鼠胚胎细胞系107合成腺病毒特异性RNA（ad-RNA），其在细胞质中作为mRNA发挥作用。我们利用ad-RNA作为mRNA代谢的模型，并且在一项初步研究中，通过与固定在滤膜上的腺病毒DNA杂交，对细胞核和细胞质中的ad-RNA进行了表征。结果表明，as-RNA在细胞核中积累，并且含有细胞质聚腺苷酸[poly(A)]的ad-RNA以数小时的半衰期周转。脉冲追踪实验证实了这些观察结果，并确定含有poly(A)的细胞质ad-RNA的半衰期约为h。第二类ad-RNA保留在细胞核中，在那里它以更长的半衰期（约24小时）周转。在37℃下，HSV对107细胞的感染受到限制，每个细胞的爆发量为5个空斑形成单位（PFU），并允许宿主DNA继续合成。感染后7小时，蛋白质合成被抑制超过50%，感染后4小时，总RNA合成被抑制50%。在感染后的前8小时内，通过对核RNA样品进行杂交测定，HSV对ad-RNA的合成速率几乎没有影响，但是，在同一时期，HSV抑制了含有poly(A)的ad-RNA在细胞质中的积累。在整个这段时间内，这种抑制程度稳步增加，在感染后6.5至8小时达到60%。未观察到对总细胞中含有poly(A)的RNA积累有显著影响。从这些实验得出的结论是，HSV感染改变了ad-RNA的代谢，从而阻止了含有poly(A)的mRNA在细胞质中的正常出现。在感染受到限制的条件下，ad-RNA的结果可能不代表总细胞中含有poly(A)的RNA的行为。
