Differential expression of the major immediate early gene of human cytomegalovirus.
Differential expression of the major immediate early gene of human cytomegalovirus.
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DOI：
10.1099/0022-1317-71-1-115
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发表时间：
1990
期刊：
影响因子：
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通讯作者：
Y. Tsutsui;T. Nogami-Satake
中科院分区：
文献类型：
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作者：
Y. Tsutsui;T. Nogami-Satake
我们制备了一种鼠单克隆抗体，可与人巨细胞病毒（HCMV）诱导的核蛋白反应，其Mr为68，000。68 K蛋白的表达进行了比较与主要立即早期（IE）72 K蛋白在各种类型的细胞感染HCMV或显微注射含有主要IE基因的质粒DNA后。在感染HCMV的人胚肺（HEL）细胞中，68 K核蛋白在72 K蛋白出现后2 ~ 3 h被检测到。68 K蛋白在感染后期主要分布于细胞质中，而72 K蛋白则主要分布于细胞核中。在IE条件下通过免疫沉淀在细胞中几乎检测不到68 K蛋白，但是，在将仅含有主要IE区域（区域1）的克隆DNA显微注射到HEL细胞的细胞核中后，68 K蛋白与72 K蛋白一起表达。72 K蛋白在注射后2 h在细胞核中表达，而68 K蛋白在注射后4 - 5 h在细胞核中表达。68 K蛋白在显微注射后在非允许的啮齿动物成纤维细胞或非允许的转化的人细胞中表达，其中这些蛋白在病毒感染后不表达。免疫沉淀后，追逐标记IE条件下或后部分digestion表明，68 K蛋白既不是降解，也不是一个主要的IE 72 K蛋白的修饰产物。
We prepared a murine monoclonal antibody reactive to a human cytomegalovirus (HCMV)-induced nuclear protein with an Mr of 68,000. Expression of the 68K protein was compared with the major immediate early (IE) 72K protein in various cell types after infection with HCMV or microinjection of plasmid DNA containing the major IE gene. The 68K nuclear protein was detected 2 to 3 h after appearance of the 72K protein in human embryonal lung (HEL) cells infected with HCMV. The 68K protein was distributed throughout the cytoplasm in the late phase of infection, while the 72K protein remained chiefly in the nucleus. The 68K protein was barely detected in the cells under IE conditions by immunoprecipitation, but, together with the 72K protein, it was expressed after microinjection of cloned DNA, containing only the major IE region (region 1), into the nuclei of HEL cells. The 72K protein was expressed in nuclei 2 h after microinjection, whereas the 68K protein was detected 4 to 5 h after the injection. The 68K protein was expressed after microinjection in non-permissive rodent fibroblasts or non-permissive transformed human cells in which these proteins were not expressed after viral infection. Immunoprecipitations after chase-labelling from IE conditions or after partial digestions suggested that the 68K protein is neither a degradation nor a modification product of the major IE 72K protein.