Burge B W, Pfefferkorn E R
J Virol. 1967 Oct;1(5):956-62. doi: 10.1128/JVI.1.5.956-962.1967.
Temperature-sensitive mutants of Sindbis virus fail to grow at a temperature that permits growth of the wild type, but when certain pairs of these mutants, mixed together, infect cells at that temperature, viral growth (i.e., complementation) occurs. The yield from this complementation, however, is of the same order of magnitude as the infectivity in the inoculum. Since in animal virus infections the protein components of the virion probably enter the cell with the viral nucleic acid, it was necessary to demonstrate that the observed complementation required synthesis of new viral protein and nucleic acid rather than some sort of rearrangement of the structural components of the inoculum. To demonstrate that complementation does require new biosynthesis, three biochemical events of normal virus growth have been observed during complementation and correlated with the efficiency of viral growth seen in complementation. These events include: (i) entrance of parental viral ribonucleic acid (RNA) into a double-stranded form; (ii) subsequent synthesis of viral RNA; and (iii) synthesis and subsequent incorporation of viral protein(s) into cell membranes where they were detected by hemadsorption. Although the infecting single-stranded RNA genome of the wild type was converted to a ribonuclease-resistant form, the genome of a mutant (ts-11) incapable of RNA synthesis at a nonpermissive temperature was not so converted. However, during complementation with another mutant also defective in viral RNA synthesis, some of the RNA of mutant ts-11 was converted to a ribonuclease-resistant form, and total synthesis of virus-specific RNA was markedly enhanced. The virus-specific alteration of the cell surface, detected by hemadsorption, was also extensively increased during complementation. These observations support the view that complementation between temperature-sensitive mutants and replication of wild-type virus are similar processes.
辛德毕斯病毒的温度敏感突变体在允许野生型生长的温度下无法生长，但当这些突变体中的某些对混合在一起，并在该温度下感染细胞时，病毒生长（即互补作用）就会发生。然而，这种互补作用产生的病毒产量与接种物中的感染性处于同一数量级。由于在动物病毒感染中，病毒粒子的蛋白质成分可能与病毒核酸一起进入细胞，因此有必要证明观察到的互补作用需要合成新的病毒蛋白质和核酸，而不是接种物结构成分的某种重排。为了证明互补作用确实需要新的生物合成过程，在互补作用过程中观察到了正常病毒生长的三个生化事件，并将其与互补作用中观察到的病毒生长效率相关联。这些事件包括：（i）亲本病毒核糖核酸（RNA）转变为双链形式；（ii）随后合成病毒RNA；以及（iii）合成病毒蛋白质并随后将其整合到细胞膜中，通过血细胞吸附检测到这些蛋白质。尽管野生型感染性单链RNA基因组转变为抗核糖核酸酶的形式，但在非允许温度下无法进行RNA合成的突变体（ts - 11）的基因组并未如此转变。然而，在与另一个在病毒RNA合成方面也有缺陷的突变体互补时，突变体ts - 11的一些RNA转变为抗核糖核酸酶的形式，并且病毒特异性RNA的总合成显著增强。在互补作用过程中，通过血细胞吸附检测到的细胞表面的病毒特异性改变也大幅增加。这些观察结果支持了温度敏感突变体之间的互补作用与野生型病毒复制是相似过程的观点。
