Ultrastructural localization of a platelet-derived growth factor/v-sis-related protein(s) in cytoplasm and nucleus of simian sarcoma virus-transformed cells.
Ultrastructural localization of a platelet-derived growth factor/v-sis-related protein(s) in cytoplasm and nucleus of simian sarcoma virus-transformed cells.
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血小板衍生生长因子/v-sis 相关蛋白在猿肉瘤病毒转化细胞的细胞质和细胞核中的超微结构定位。
DOI：
10.1073/pnas.84.8.2317
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发表时间：
1987
影响因子：
11.1
通讯作者：
Deuel,TF
中科院分区：
文献类型：
--
作者：
Yeh,HJ;Pierce,GF;Deuel,TF
应用兔抗血小板源性生长因子（PDGF）多克隆抗血清，用免疫荧光和蛋白A金标记技术，分析了三种猴肉瘤病毒（SSV）转化细胞系中v-sis相关蛋白的亚细胞分布。抗原反应蛋白在与蛋白质合成和加工相关的亚细胞器中被识别，包括多聚核糖体、内质网和高尔基体，以及质膜的细胞质表面。使用Lowicryl K4 M树脂包埋技术，在完整细胞和分离的细胞核中也显示出与核染色质相关的突出免疫反应蛋白。如果在包埋过程中用Epon树脂代替Lowicryl或如果切片用不相关的抗血清，则在与抗PDGF孵育的非SSV转化的成纤维细胞切片中蛋白A-金标记显著减少，并且在SSV转化的细胞中不存在。使用从SSV转化的成纤维细胞分离的细胞核，在基于硝化纤维素的免疫测定中证实了v-sis相关抗原的核定位。因此，抗原性识别为与v-sis基因产物相关的多肽不仅可以在与蛋白质合成和包装相关的亚细胞器中发现，而且可以在SSV转化细胞的细胞核中发现。这些结果提高了可能性，v-sis-或PDGF-样蛋白可能在细胞核内的SSV转化细胞的功能。
The subcellular distribution of v-sis-related protein(s) was analyzed in three simian sarcoma virus (SSV)-transformed cell lines with immunofluorescence and protein A-gold labeling techniques using rabbit polyclonal anti-platelet-derived-growth factor (PDGF) antisera. Antigenically reactive proteins were recognized in subcellular organelles related to protein synthesis and processing, including polyribosomes, endoplasmic reticulum, and the Golgi apparatus, as well as on the cytoplasmic surface of plasma membranes. Prominent immunoreactive proteins were also shown in association with nuclear chromatin in intact cells and in isolated nuclei using Lowicryl K4M resin embedding techniques. Protein A-gold labeling was markedly reduced in sections of non-SSV-transformed fibroblasts incubated with anti-PDGF and absent from SSV-transformed cells if Epon resin was substituted for Lowicryl in the embedding process or if sections were with irrelevant antisera. Nuclear localization of v-sis-related antigens was confirmed in a nitrocellulose-based immunoassay using nuclei isolated from SSV-transformed fibroblasts. Thus, polypeptides recognized antigenically as related to the v-sis gene product not only may be found in subcellular organelles associated with protein synthesis and packaging but also may be found in the nucleus of SSV-transformed cells. These results raise the possibility that v-sis- or PDGF-like proteins may function within the nucleus of SSV-transformed cells.