Yager D R, Bachenheimer S L
Department of Microbiology and Immunology, University of North Carolina, Chapel Hill 27599.
Virus Genes. 1988 Mar;1(2):135-48. doi: 10.1007/BF00555933.
The effects of productive herpes simplex virus infection on host gene expression were examined by measuring the rates of synthesis and subsequent fates of several Vero cell mRNAs. The rates of transcription of actin, beta-tubulin, and histone-3 and -4 RNAs were measured by pulse-labeling of intact cells as well as by run-on transcription in isolated nuclei. At both early (2 hr) and late (6 hr) times, the relative rates of transcription of these RNAs were greater than in uninfected cells. Kinetic labeling experiments performed at late times also revealed increased turnover rates of nuclear RNAs. That the rate of appearance of these RNAs in the cytoplasm was also reduced suggests that these cellular RNAs are being specifically retained and degraded in the nucleus. Levels of pre-existing cytoplasmic RNAs as measured by Northern blot analysis declined rapidly after infection though the nuclear steady-state levels of these RNAs increased up to 3 hr postinfection and then declined between 3 and 10 hr postinfection. At no time was the accumulation of processing intermediates detectable. Finally, we also determined that, consistent with the decline in levels of histone mRNA, rates of histone protein synthesis declined rapidly after infection.
通过测量几种非洲绿猴肾细胞（Vero细胞）mRNA的合成速率及其后续去向，研究了单纯疱疹病毒感染对宿主基因表达的影响。通过对完整细胞进行脉冲标记以及在分离的细胞核中进行连续转录，测量肌动蛋白、β-微管蛋白以及组蛋白H3和H4 RNA的转录速率。在早期（2小时）和晚期（6小时），这些RNA的相对转录速率均高于未感染细胞。在晚期进行的动力学标记实验还显示，核RNA的周转率增加。这些RNA在细胞质中出现的速率也降低，这表明这些细胞RNA在细胞核中被特异性保留并降解。通过Northern印迹分析测量，感染前存在的细胞质RNA水平在感染后迅速下降，尽管这些RNA的核稳态水平在感染后3小时内升高，然后在感染后3至10小时之间下降。在任何时候都未检测到加工中间体的积累。最后，我们还确定，与组蛋白mRNA水平下降一致，感染后组蛋白的合成速率迅速下降。
