Differential expression and distinct structure of 69- and 100-kDa forms of 2-5A synthetase in human cells treated with interferon.
Differential expression and distinct structure of 69- and 100-kDa forms of 2-5A synthetase in human cells treated with interferon.
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DOI：
10.1016/s0021-9258(17)44794-6
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发表时间：
1990-10
期刊：
影响因子：
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通讯作者：
I. Marié;J. Svab;N. Robert;J. Galabru;A. Hovanessian
中科院分区：
文献类型：
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作者：
I. Marié;J. Svab;N. Robert;J. Galabru;A. Hovanessian
使用特异性单克隆和多克隆抗体，在α-干扰素处理的人HeLa和Daudi细胞中研究了69-和100-kDa形式的2 '，5'-寡腺苷酸（2-5A）合成酶（p69和p100）的诱导、合成和亚细胞定位。虽然p69和p100都是由干扰素诱导的，但在这些细胞系中，干扰素的剂量反应和每种蛋白质的合成动力学存在显著差异。这两种蛋白质主要定位于细胞质中。然而，干扰素处理的细胞的免疫酶染色分析表明，一部分p69是集中在细胞核周围，其余的是分布在一个特定的模式在细胞质中，而p100被发现在细胞质中的扩散状态。与其与细胞膜的结合雅阁，p69被豆蔻酰化。[35 S]半胱氨酸和[3 H]肉豆蔻酸标记的p69制剂通过二维凝胶等电聚焦分离，pI值分别在7.0-8.0和7.0-7.5范围内。这些观察结果表明存在两种形式的p69，它们可能作为二聚体复合在一起。有利于p69的二聚体形式，在凝胶过滤实验中，p69的峰通常在对应于160 kDa分子量的级分中发现，而p100作为单体回收。综上所述，这些结果表明，p69和p100具有不同的特征，它们的表达是每种细胞类型对干扰素应答的特异性。
Using specific monoclonal and polyclonal antibodies, the induction, synthesis and subcellular localization of 69- and 100-kDa forms of 2‘,5‘-oligoadenylate (2-5A) synthetase (p69 and p100) were investigated in alpha-interferon-treated human HeLa and Daudi cells. Although both p69 and p100 were induced by interferon, there were significant differences in the interferon dose-response and the kinetics of synthesis of each protein in these cell lines. Both proteins are localized mainly in the cytoplasm. However, immunoenzymatic staining analysis of interferon-treated cells indicated that a proportion of p69 is concentrated around the nuclei and the rest is distributed in a specific pattern in the cytoplasm whereas p100 is found in a diffuse state in the cytoplasm. In accord with its association to cell membranes, p69 is myristilated. The [35S]cysteine and [3H]myristate-labeled p69 preparations were resolved by two-dimensional gel isoelectric focusing with pI values in the pH range of 7.0-8.0 and 7.0-7.5, respectively. These observations suggested the existence of two forms of p69 which might be complexed together as a dimer. In favor with a dimeric form of p69, in gel filtration experiments the peak of p69 was routinely found in fractions corresponding to a molecular mass of 160 kDa whereas p100 was recovered as a monomer. Taken together, these results indicate that p69 and p100 have distinct characteristics and that their expression is a specific property of each cell type in response to interferon.