Identification of homo- and heteromeric interactions between members of the breast carcinoma-associated D52 protein family using the yeast two-hybrid system
Identification of homo- and heteromeric interactions between members of the breast carcinoma-associated D52 protein family using the yeast two-hybrid system
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DOI：
10.1038/sj.onc.1201604
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发表时间：
1998-02-19
期刊：
影响因子：
8
通讯作者：
Gunning, P
中科院分区：
文献类型：
--
作者：
Byrne, JA;Nourse, CR;Gunning, P
hD52基因最初是通过其在人类乳腺癌中的表达水平升高而确定的。从其他物种中克隆的D52同源物表明，D52可能在钙介导的信号转导和细胞增殖中发挥作用。hD52的两个人类同源物hD53和hD54也已被鉴定出来，这表明存在一个新的基因/蛋白家族。由于d52样蛋白序列都被预测含有一个螺旋结构域，我们使用酵母双杂交系统和谷胱甘肽s -转移酶下拉试验来研究d52样蛋白之间是否存在同源和/或异源相互作用。对共转染配对D52-like结构体的酵母菌株的分析表明，D52-like融合蛋白通过其预测的卷曲结构域以同源和异源方式相互作用。同样，对人类乳腺癌表达文库的广泛双杂交筛选发现，hD53和hD52是hD52和hD53诱饵的潜在相互作用物。因此，d52样蛋白似乎通过与其他d52样蛋白的特定相互作用来发挥和/或调节其活性，这反过来可能是这些分子在控制细胞增殖中的潜在作用所固有的。
The hD52 gene was originally identified through its elevated expression level in human breast carcinoma. Cloning of D52 homologues from other species has indicated that D52 may play roles in calcium-mediated signal transduction and cell proliferation. Two human homologues of hD52, hD53 and hD54, have also been identified, demonstrating the existence of a novel gene/protein family. Since D52-like protein sequences are all predicted to contain a coiled-coil domain, we used the yeast two-hybrid system and glutathione S-transferase pull-down assays to investigate whether homo- and/or heteromeric interactions occur between D52-like proteins. Analyses of yeast strains co-transfected with paired D52-like constructs indicated that D52-like fusion proteins interact in homo- and heteromeric fashions through their predicted coiled-coil domains. Similarly, extensive two-hybrid screenings of a human breast carcinoma expression library identified hD53 and hD52 as potential interactors for both hD52 and hD53 baits. Thus, D52-like proteins appear to exert and/or regulate their activities through specific interactions with other D52-like proteins, which in turn may be intrinsic to potential roles of these molecules in controlling cell proliferation.