Buchwald I, Bialdiga M, Traub U, Traub P
Mol Biol Rep. 1978 Feb 28;4(1):9-13. doi: 10.1007/BF00775173.
The distribution of poly(A)+ mRNA among polysomes, monosomes, and ribosome-free supernatant fractions after mengovirus infection of Ehrlich ascites tumor (EAT) cells was investigated employing sucrose gradient centrifugation of their corresponding postnuclear supernatants. Poly(A)+ mRNA was isolated from sucrose gradient fractions and quantitated in a cell-free protein synthesizing system from uninfected EAT cells. It was also localized by annealing [3H]-poly(U) to the poly(A)-tracts of mRNA present in the sucrose gradient fractions. Both experiments revealed a gradual shift of host poly(A)+ mRNA from large to small polysomes and monosomes, respectively, with the time postinfection. The greatest part of host template RNA appears to remain ribosome-bound and only a fraction seems to be detached from the ribosomes in the course of mengovirus infection. At the end of the infectious cycle, 8 h postinfection, approximately 70% of the poly(A)+ mRNA detected in uninfected cells is still biologically active, but not translated in vivo, in agreement with data from the [3H]poly(U)hybridization experiment.
采用对艾氏腹水瘤(EAT)细胞的相应核后上清液进行蔗糖梯度离心的方法,研究了脑心肌炎病毒感染EAT细胞后,多聚腺苷酸(poly(A))+信使核糖核酸(mRNA)在多核糖体、单核糖体和无核糖体上清液组分中的分布情况。从蔗糖梯度级分中分离出poly(A)+ mRNA,并在来自未感染EAT细胞的无细胞蛋白质合成系统中进行定量分析。还通过将[3H] - 聚尿苷酸(poly(U))与蔗糖梯度级分中存在的mRNA的poly(A)尾退火,对其进行定位。两项实验均显示,感染后随着时间的推移,宿主poly(A)+ mRNA分别从大多核糖体逐渐转移至小多核糖体和单核糖体。在脑心肌炎病毒感染过程中,宿主模板RNA的大部分似乎仍与核糖体结合,只有一小部分似乎从核糖体上脱离。在感染周期结束时,即感染后8小时,未感染细胞中检测到的约70%的poly(A)+ mRNA在生物学上仍然具有活性,但在体内未被翻译,这与[3H]poly(U)杂交实验的数据一致。