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氧还蛋白Ref1 是一种双功能蛋白质, 在细胞氧还调控和DNA 无嘌呤/无嘧啶损伤修复中起重要作用。为寻找与它相互作用的蛋白Rips(Ref1 interacting proteins) , 用Ref1 氧还功能域进行了酵母双杂交库的筛选, 得到了5 种阳性克隆。其中Rip3 经测序证实为泛蛋白连接酶Ubc9。Hela 细胞中共过表达Ubc9 可以明显抑制Ref1对AP1 报告系统的增强作用。Western 印迹发现Ref1 的蛋白质水平由于Ubc9 的共过表达而下调。这些结果表明Ubc9 参与了Ref1 的蛋白质降解, 从而下调了Ref1 的生理功能。
Ref 1 is a bifunctional protein that plays an important role in the regulation of cellular oxygenation and DNA apathetic / apyrimidinic damage repair. In order to find the protein Rips interacting with it, we screened yeast two-hybrid library with Ref 1 oxygen functional domain and got five positive clones. Among them, Rip3 was confirmed to be ubiquitin ligase Ubc9 by sequencing. The overexpression of Ubc9 in Hela cells can significantly inhibit the enhancement of Ref1 on AP1 reporter system. Western blotting revealed that the protein level of Ref-1 was down-regulated due to the co-overexpression of Ubc9. These results indicate that Ubc9 is involved in the protein degradation of Ref-1 and thus down-regulates the physiological function of Ref-1.