Stinski M F
J Virol. 1977 Sep;23(3):751-67. doi: 10.1128/JVI.23.3.751-767.1977.
In cytomegalovirus-infected cells, the rate of protein synthesis was detected as two peaks. One occurred during the early phase of infection, 0 to 36 h postinfection, and the other occurred during the late phase, after the initiation of viral DNA synthesis. Double-isotopic-label difference analysis demonstrated that host and viral proteins were synthesized simultaneously during both phases. In the early phase, approximately 70 to 90% of the total proteins synthesized were host proteins, whereas approximately 10 to 30% were viral, even at a multiplicity of infection of 20 PFU/cell. Virus-related proteins or glycoproteins were referred to as infected-cell specific (ICS). Two ICS glycoproteins (gp145 and 100) were clearly detectable and were synthesized preferentially in the early phase of infection. Their synthesis was concomitant with stimulation of the protein synthesis rate. In the late phase of infection, approximately 50 to 60% of the total protein synthesis was viral and approximately 40 to 50% was host. The ICS proteins and glycoproteins detected during the late phase of infection were viral structural proteins. Infectious virus was not detectable until 48 to 72 h postinfection. An inhibitor of viral DNA synthesis, phosphonoacetic acid, prevented the appearance of the late-phase ICS proteins and glycoproteins, but there was little or no effect on early ICS glycoprotein synthesis. Radiolabeled ICS proteins and glycoproteins were identified by their relative rates of synthesis, by their different electrophoretic mobilities compared with those of host proteins and host glycoproteins, and by their similar electrophoretic mobilities compared to those of proteins and glycoproteins associated with virions and dense bodies of cytomegalovirus. Structural viral antigens in the infected-cell extracts were removed by immunoprecipitation, using F(ab')(2) fragments of cytomegalovirus-specific antibodies, and identified as described above. The last two criteria were used to identify viral structural ICS proteins and glycoproteins. Although approximately 35 structural proteins were found to be associated with purified virions and dense bodies, the continued synthesis of host cell proteins complicated their identification in infected cells. Nevertheless, seven of the nine structural glycoproteins were identified as ICS glycoproteins.
在巨细胞病毒感染的细胞中，蛋白质合成速率检测到有两个峰值。一个出现在感染的早期阶段，即感染后0至36小时，另一个出现在晚期阶段，即在病毒DNA合成开始之后。双同位素标记差异分析表明，宿主蛋白和病毒蛋白在两个阶段均同时合成。在早期阶段，即使感染复数为20 PFU/细胞，合成的总蛋白中约70%至90%是宿主蛋白，而约10%至30%是病毒蛋白。与病毒相关的蛋白或糖蛋白被称为感染细胞特异性（ICS）蛋白。两种ICS糖蛋白（gp145和100）清晰可检测到，且在感染早期优先合成。它们的合成与蛋白质合成速率的刺激同时发生。在感染晚期，总蛋白合成中约50%至60%是病毒蛋白，约40%至50%是宿主蛋白。在感染晚期检测到的ICS蛋白和糖蛋白是病毒结构蛋白。直到感染后48至72小时才能检测到感染性病毒。病毒DNA合成抑制剂膦甲酸可阻止晚期ICS蛋白和糖蛋白的出现，但对早期ICS糖蛋白合成几乎没有影响。通过合成的相对速率、与宿主蛋白和宿主糖蛋白相比不同的电泳迁移率以及与巨细胞病毒病毒体和致密体相关的蛋白和糖蛋白相比相似的电泳迁移率来鉴定放射性标记的ICS蛋白和糖蛋白。使用巨细胞病毒特异性抗体的F(ab')(2)片段通过免疫沉淀去除感染细胞提取物中的结构病毒抗原，并按上述方法进行鉴定。最后两个标准用于鉴定病毒结构ICS蛋白和糖蛋白。尽管发现约35种结构蛋白与纯化的病毒体和致密体相关，但宿主细胞蛋白的持续合成使其在感染细胞中的鉴定变得复杂。然而，9种结构糖蛋白中有7种被鉴定为ICS糖蛋白。
