LOCALIZATION OF THE E1B PROTEINS OF ADENOVIRUS-5 IN TRANSFORMED-CELLS, AS REVEALED BY INTERACTION WITH MONOCLONAL-ANTIBODIES
LOCALIZATION OF THE E1B PROTEINS OF ADENOVIRUS-5 IN TRANSFORMED-CELLS, AS REVEALED BY INTERACTION WITH MONOCLONAL-ANTIBODIES
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DOI：
10.1016/0042-6822(85)90421-0
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发表时间：
1985-01-01
期刊：
影响因子：
3.7
通讯作者：
VANDEREB, AJ
中科院分区：
文献类型：
--
作者：
ZANTEMA, A;FRANSEN, JAM;VANDEREB, AJ
单克隆抗体（一种针对 5 型腺病毒 E1B 55-kDa [千道尔顿] 蛋白，另一种针对 E1B 21-kDa 蛋白）已被分离出来，并显示可识别与抗肿瘤血清相同的蛋白。使用这些单克隆抗体对含有完整腺病毒早期区域 1 的转化大鼠肾细胞进行的免疫荧光研究表明，E1B 21-kDa 蛋白位于核周区域。 E1B 55-kDa 蛋白定位在许多不同的位点：在靠近细胞核的细胞质中的离散体中观察到强荧光，在细胞与细胞接触处观察到中等荧光，在细胞质中观察到弱染色。与 E1B 55-kDa 蛋白相关的细胞 p53 抗原也存在于离散的细胞质体中，但在细胞与细胞接触处没有或仅少量存在。但离散体以外的细胞质中未见p53，但细胞核呈弱阳性。通过 EM 进一步研究了离散细胞质体的性质，发现其由一簇 8 nm 丝组成。丝的直径与细胞骨架中间丝的直径相似。用针对各种中间丝蛋白的抗体染色没有显示出与簇的显着反应，而波形蛋白中间丝可以在细胞中以典型的细胞骨架模式被证明。还表明该簇不是由错误聚集的微管蛋白组成的。
Monoclonal antibodies, one against the adenovirus type 5 E1B 55-kDa [kilodalton] protein and one against the E1B 21-kDa protein, have been isolated and shown to recognize the same proteins as antitumor sera. Immunofluorescence studies with these monoclonal antibodies on transformed rat kidney cells containing the complete adenovirus early region 1, showed that the E1B 21-kDa protein is localized in the perinuclear region. The E1B 55-kDa protein is localized in a number of different sites: a strong fluorescence is observed in a discrete body in the cytoplasm close to the nucleus, a moderate fluorescence is found in cell-cell contacts and a weak staining in the cytoplasm. The cellular p53 antigen, which is associated with the E1B 55-kDa protein, is also found in the discrete cytoplasmic body, but not, or only in small amounts at the cell-cell contacts. However, p53 is not seen in the cytoplasm outside the discrete body, but the nucleus is weakly positive. The nature of the discrete cytoplasmic body was investigated further by EM and was found to be composed to a cluster of 8-nm filaments. The diameter of the filaments is similar to that of cytoskeletal intermediate filaments. Staining with antibodies against the various intermediate filament proteins did not show a significant reaction with the cluster, while vimentin intermediate filaments could be demonstrated in the cells in a typical cytoskeletal pattern. It was also shown that the cluster is not composed of incorrectly aggregated tubulin.