SYNTHESIS OF PROTEINS AND GLYCOPROTEINS IN CELLS INFECTED WITH HUMAN CYTOMEGALOVIRUS
SYNTHESIS OF PROTEINS AND GLYCOPROTEINS IN CELLS INFECTED WITH HUMAN CYTOMEGALOVIRUS
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DOI：
10.1128/jvi.23.3.751-767.1977
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发表时间：
1977-01-01
影响因子：
5.4
通讯作者：
STINSKI, MF
中科院分区：
文献类型：
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作者：
STINSKI, MF
在巨细胞病毒感染的[人成纤维细胞]细胞中，检测到蛋白质合成率为2个峰。一个发生在感染早期，感染后0-36小时，另一个发生在后期，在病毒DNA合成开始后。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同时合成。在早期阶段，合成的总蛋白中约有70-90%是宿主蛋白，但约有10-30%是病毒蛋白，即使在20个PFU（斑块形成单位）/细胞的感染倍数下也是如此。病毒相关蛋白或糖蛋白被称为感染细胞特异性（ICS）。两种ICS糖蛋白（gp145和100）可明显检测到，并在感染早期优先合成。它们的合成伴随着蛋白质合成速率的刺激。在感染后期，总蛋白质合成中约有50-60%是病毒，约有40-50%是宿主。感染后期检测到的ICS蛋白和糖蛋白为病毒结构蛋白。感染后48-72小时才检出感染病毒。一种病毒DNA合成抑制剂磷酸乙酸可以阻止后期ICS蛋白和糖蛋白的出现，但对早期ICS糖蛋白的合成几乎没有影响。放射性标记的ICS蛋白和糖蛋白通过它们的相对合成速率、与宿主蛋白和宿主糖蛋白不同的电泳迁移率以及与病毒粒子和巨细胞病毒致密体相关的蛋白和糖蛋白相似的电泳迁移率来鉴定。利用巨细胞病毒特异性抗体F（ab”）2片段，免疫沉淀法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2项标准用于鉴定病毒结构ICS蛋白和糖蛋白。虽然大约有35种结构蛋白与纯化的病毒粒子和致密体相关，但宿主细胞蛋白的持续合成使它们在感染细胞中的鉴定变得复杂。然而，9个结构糖蛋白中有7个被鉴定为ICS糖蛋白。
In cytomegalovirus-infected [human fibroblast] cells, the rate of protein synthesis was detected as 2 peaks. One occurred during the early phase of infection, 0-36 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, about 70-90% of the total proteins synthesized were host proteins, but about 10-30% were viral, even at a multiplicity of infection of 20 PFU[plaque forming units]/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, about 50-60% of the total protein synthesis was viral and about 40-50% was host. The ICS proteins and glycoproteins detected during the late phase of infection were viral structural proteins. Infectious virus was not detectable until 48-72 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 2 criteria were used to identify viral structural ICS proteins and glycoproteins. Although about 35 structural proteins were associated with purified virions and dense bodies, the continued synthesis of host cell proteins complicated their identification in infected cells. Nevertheless, 7 of the 9 structural glycoproteins were identified as ICS glycoproteins.