Lemieux R, Vassef A, Ben-Hamida F, Beaud G
Eur J Biochem. 1982 Dec 15;129(2):265-71. doi: 10.1111/j.1432-1033.1982.tb07048.x.
Translation of cellular and early vaccinia RNA in nuclease-treated lysates, derived from uninfected and vaccinia-virus-infected cells at the early stage, has been investigated. When using limiting amounts of RNA no discrimination of translation was observed in the infected cells lysates; this conclusion was confirmed by sensitive RNA competition experiments for translation in vitro and also when using two different fractionated systems for protein synthesis in vitro. This absence of detectable discrimination in vitro was established both by comparing incorporation of [35S]methionine into proteins and by analysis of the products thus synthesized by sodium dodecylsulfate gel electrophoresis. However, a modification of the translational machinery from vaccinia-virus-infected cells did occur since the only the ribosomal salt wash derived from infected cells was able to reverse the inhibition of protein synthesis in vitro resulting from excess RNA (control or early). This property of vaccinia-virus-infected cell lysates may result from the synthesis l machinery from vaccinia-virus-infected cells did occur since the only the ribosomal salt wash derived from infected cells was able to reverse the inhibition of protein synthesis in vitro resulting from excess RNA (control or early). This property of vaccinia-virus-infected cell lysates may result from the synthesis l machinery from vaccinia-virus-infected cells did occur since the only the ribosomal salt wash derived from infected cells was able to reverse the inhibition of protein synthesis in vitro resulting from excess RNA (control or early). This property of vaccinia-virus-infected cell lysates may result from the synthesis of an early protein involved in translation or from a better recovery of translational factors from the infected cells, as suggested in the accompanying paper.
对来自未感染细胞和早期牛痘病毒感染细胞的经核酸酶处理的裂解物中的细胞和早期牛痘病毒RNA的翻译进行了研究。当使用限量RNA时，在感染细胞裂解物中未观察到翻译的差异；这一结论通过体外翻译的灵敏RNA竞争实验得到证实，并且在使用两种不同的体外蛋白质合成分级系统时也得到了证实。通过比较[35S]甲硫氨酸掺入蛋白质的情况以及通过十二烷基硫酸钠凝胶电泳分析由此合成的产物，确定了体外不存在可检测到的差异。然而，牛痘病毒感染细胞的翻译机制确实发生了改变，因为只有来自感染细胞的核糖体盐洗能够逆转由过量RNA（对照或早期）导致的体外蛋白质合成抑制。牛痘病毒感染细胞裂解物的这一特性可能是由于合成了一种参与翻译的早期蛋白质，或者如随附论文中所指出的，是由于从感染细胞中更好地回收了翻译因子。
