Baines J D, Jacob R J, Simmerman L, Roizman B
Marjorie B. Kovler Viral Oncology Laboratories, University of Chicago, Illinois 60637.
J Virol. 1995 Feb;69(2):825-33. doi: 10.1128/JVI.69.2.825-833.1995.
Earlier studies have shown that the UL11 gene of herpes simplex virus encodes a myristylated virion protein and that the UL11 gene enables efficient virion envelopment and export from infected cells. A rabbit polyclonal antibody directed against an affinity-purified UL11-glutathione-S-transferase fusion protein was made and used to study the properties of the UL11 protein and its distribution in infected cells. We report the following: (i) UL11 protein formed up to five bands (apparent M(r)s, 17,000 to 22,000) in denaturing polyacrylamide gels; (ii) fluorescent-antibody studies revealed the presence of UL11 protein in the perinuclear space and in sites within the nucleus; (iii) immune electron microscopic studies indicated that the UL11 gene products were associated with the inner nuclear membrane, with cytoplasmic membranes and ribbon-like cytoplasmic structures resembling membranous organelles, with nuclear bodies shown by fluorescence microscopy to be different from nucleoli in which US11 protein accumulates, and with enveloped virions but not with nuclear capsids; and (iv) the nuclear bodies containing UL11 protein were reminiscent both of type IV morphotypes consisting of an electron-dense core containing the UL11 proteins surrounded by a more electron-transluscent core and of type V morphotypes consisting of material homogenous in electron opacity. We conclude that (i) the UL11 protein is processed after synthesis; (ii) the localization of UL11 protein with virions and membranes is consistent with the hypothesis that UL11 plays a role in the transport of virions to the extracellular space; and (iii) although the significance of the association of UL11 proteins with nuclear bodies is unknown, the results indicate that nuclear bodies differ with respect to their morphologies and contents of viral protein and suggest that UL11 protein may have more than one function in the infected cell.
早期研究表明,单纯疱疹病毒的UL11基因编码一种肉豆蔻酰化的病毒体蛋白,且UL11基因能使病毒体有效地被包裹并从受感染细胞中输出。制备了一种针对亲和纯化的UL11-谷胱甘肽-S-转移酶融合蛋白的兔多克隆抗体,并用于研究UL11蛋白的特性及其在受感染细胞中的分布。我们报告如下:(i) 在变性聚丙烯酰胺凝胶中,UL11蛋白形成多达五条带(表观分子量为17,000至22,000);(ii) 荧光抗体研究显示,UL11蛋白存在于核周间隙和细胞核内的位点;(iii) 免疫电子显微镜研究表明,UL11基因产物与内核膜、细胞质膜以及类似于膜性细胞器的带状细胞质结构相关,荧光显微镜显示核体与US11蛋白积累的核仁不同,且与包膜病毒体相关,但与核衣壳无关;(iv) 含有UL11蛋白的核体让人联想到IV型形态型,即由一个含有UL11蛋白的电子致密核心被一个电子透明度更高的核心包围,以及V型形态型,即由电子不透明度均匀的物质组成。我们得出以下结论:(i) UL11蛋白在合成后会被加工;(ii) UL11蛋白与病毒体和膜的定位与UL11在病毒体向细胞外空间运输中起作用的假设一致;(iii) 尽管UL11蛋白与核体关联的意义尚不清楚,但结果表明核体在其形态和病毒蛋白含量方面存在差异,并提示UL11蛋白在受感染细胞中可能具有不止一种功能。