Fischinger P F, Tuttle-Fuller N, Hüper G, Bolognesi D P
J Virol. 1975 Aug;16(2):267-74. doi: 10.1128/JVI.16.2.267-274.1975.
Cloned 3T3FL cells were synchronized in G1 phase of the cell cycle by deprivation of multiplication stimulatory activity of serum and were then infected with Moloney leukemia virus. Eclipse period of virus could be made to vary from less than 10 to 34 h. All virus release was completely dependent and occurred immediately after the first mitosis following serum reconstitution. Virus yield was not affected by the time of virus inoculation as related to the cell DNA synthetic phase. Colchicine arrested the cells in mitosis and prevented the formation of infectious virus. Viral proteins p10, p30, and gp71 were assayed in cell lysates during the growth curve of virus in synchronized cells. The group-specific determinants of each protein were measured in a competition radioimmunoassay. None of the virus proteins appeared during the eclipse period of the virus. All three proteins appeared simultaneously, coincident with mitosis, and continued to rise during the G1 phase. The absolute quantities of each protein were proportional to the amount of Moloney leukemia virus produced. The relative amounts of some of the viral proteins in the cell did not correspond to their content in purified virions suggesting several possible mechanisms of control.
通过剥夺血清的增殖刺激活性,将克隆的3T3FL细胞同步于细胞周期的G1期,然后用莫洛尼白血病病毒感染。病毒的隐蔽期可从少于10小时变化到34小时。所有病毒释放完全依赖于血清重构后的第一次有丝分裂,并在其后立即发生。病毒产量不受与细胞DNA合成期相关的病毒接种时间的影响。秋水仙碱使细胞停滞在有丝分裂期,并阻止感染性病毒的形成。在同步化细胞中病毒生长曲线期间,对细胞裂解物中的病毒蛋白p10、p30和gp71进行了检测。在竞争放射免疫测定中测量了每种蛋白质的群特异性决定簇。在病毒隐蔽期没有出现任何病毒蛋白。所有三种蛋白质同时出现,与有丝分裂同时发生,并在G1期持续上升。每种蛋白质的绝对量与产生的莫洛尼白血病病毒量成比例。细胞中一些病毒蛋白的相对量与其在纯化病毒粒子中的含量不对应,提示了几种可能的调控机制。