The BPV-1 E5 oncoprotein expressed in Schizosaccharomyces pombe exhibits normal biochemical properties and binds to the endogenous 16-kDa component of the vacuolar proton-ATPase.
The BPV-1 E5 oncoprotein expressed in Schizosaccharomyces pombe exhibits normal biochemical properties and binds to the endogenous 16-kDa component of the vacuolar proton-ATPase.
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粟酒裂殖酵母中表达的 BPV-1 E5 癌蛋白表现出正常的生化特性，并与液泡质子 ATP 酶的内源 16 kDa 成分结合。
DOI：
10.1016/0042-6822(92)90932-f
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发表时间：
1992
期刊：
影响因子：
3.7
通讯作者：
Schlegel,R
中科院分区：
文献类型：
--
作者：
Goldstein,DJ;Toyama,R;Dhar,R;Schlegel,R
牛乳头瘤病毒1型的44氨基酸E5癌蛋白转化永生化小鼠成纤维细胞系。这种高度疏水的蛋白形成同型二聚体，定位于细胞膜间室（包括高尔基体），并与液泡质子atp酶的16kda膜嵌入成分（16k）形成复合物。为了建立E5/16k相互作用的遗传和生化分析系统，将E5基因克隆到裂变酵母schizosaccharomyces pombe中的表达载体中。该系统合成的E5蛋白正常二聚并与内源和过表达的ds结合。pombe16k蛋白质。这些的比较。pombebe和哺乳动物的16k蛋白在羧基末端氨基酸上表现出很强的保守性，但在氨基末端序列上有较大的差异，这表明E5与16k羧基结构域相互作用。最后，描述了一个新的蛋白表位标签，该标签首次允许E5与针对16k蛋白的抗体共沉淀。
The 44-amino-acid E5 oncoprotein of bovine papillomavirus type 1 transforms immortalized murine fibroblast cell lines. This highly hydrophobic protein forms homodimers, localizes to intracellular membrane compartments (including the Golgi apparatus), and forms a complex with the 16-kDa membrane-embedded constituent (16k) of the vacuolar proton-ATPase. To develop a system for the genetic and biochemical analysis of the E5/16k interaction, the E5 gene was cloned into a new vector which was designed for expression in the fission yeastSchizosaccharomyces pombe. The E5 protein synthesized in this system dimerized normally and bound to endogenous and overexpressedS. pombe16k protein. Comparison of theS. pombeand mammalian 16k proteins showed strong conservation in carboxyl-terminal amino acids but greater variation in the amino-terminal sequences, suggesting that E5 was interacting with the 16k carboxyl domains. Finally, a new protein epitope tag is described which permitted for the first time the coprecipitation of E5 with antibodies directed against the 16k protein.