Immunological characterization of an early cytomegalovirus single-strand DNA-binding protein with similarities to the HSV major DNA-binding protein.
Immunological characterization of an early cytomegalovirus single-strand DNA-binding protein with similarities to the HSV major DNA-binding protein.
复制标题
与 HSV 主要 DNA 结合蛋白相似的早期巨细胞病毒单链 DNA 结合蛋白的免疫学特征。
DOI：
10.1016/0042-6822(87)90154-1
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发表时间：
1987
期刊：
影响因子：
3.7
通讯作者：
Gibson,W
中科院分区：
文献类型：
--
作者：
Anders,DG;Kidd,JR;Gibson,W
针对科尔本株巨细胞病毒（CMV）的129 kDa早期单链DNA结合蛋白（DB 129）制备单特异性多克隆抗血清，并用于研究其在感染细胞中的分布及其与拟议的人CMV（HCMV）对应物（DB 140）的相关性。间接免疫荧光固定，感染的人成纤维细胞显示DB 129被定位在复制CMV的核内包涵体的特征。用每毫升50至100 μg膦甲酸处理感染的细胞导致DB 129的过量产生及其在核内的积累，包括内含物内部和核质的周围区域，而用500 μg/ml处理防止内含物形成，并且DB 129定位在整个感染细胞核的离散点。在间接免疫测定中，血清与HCMV DB 140的交叉反应估计为10%，并且它们在免疫荧光中的使用将DB 140定位于HCMV感染细胞的核内含物。它们的免疫交叉反应性，以及它们相似的生化特性和细胞内分布，支持DB 129和DB 140是同源基因的蛋白产物的可能性。这些蛋白质的关系，单纯疱疹病毒主要的DNA结合蛋白进行了讨论。
Monospecific polyclonal antisera were prepared against the 129-kDa, early, single-strand DNA-binding protein (DB129) of strain Colburn cytomegalovirus (CMV), and used to study its distribution in infected cells and its relatedness to a proposed human CMV (HCMV) counterpart (DB140). Indirect immunofluorescence of fixed, infected human fibroblasts showed DB129 to be localized within the intranuclear inclusions characteristic of replicating CMV. Treatment of infected cells with 50 to 100 μg phosphonoformic acid per milliliter resulted in the overproduction of DB129 and its accumulation within nuclei, both inside the inclusions and in surrounding areas of the nucleoplasm, whereas treatment with 500 μg/ml prevented inclusion formation, and DB129 was localized at discrete points throughout the infected-cell nuclei. The sera cross-reacted an estimated 10% with HCMV DB140 in an indirect immunoassay, and their use in immunofluorescence localized DB140 to the nuclear inclusions of HCMV-infected cells. Their immunological cross-reactivity, as well as their similar biochemical properties and intracellular distribution, support the likelihood that DB129 and DB140 are the protein products of homologous genes. The relationship of these proteins to the herpes simplex major DNA-binding protein is discussed.