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溴区包含蛋白7(BRD7)是采用cDNA代表性差异分析方法克隆的一个新基因.研究证实了BRD7能够与乙酰化的组蛋白3结合,其识别位点在组蛋白3的14位氨基酸;并且证实了溴区结构域(Bromodomain)缺失型的BRD7突变体失去了与乙酰化组蛋白3的结合能力.Bromodomain是在进化上高度保守的功能结构域,该结构域在空间构象上具有鲜明的特征:包括4个左手、呈反向平行排列的“螺旋(αz,αA,αB,αC)”以及2个“连结环”(ZAloop,BCloop).通过生物信息学等综合分析,预测BRD7可能具有上述特征.依据上述分析结果,构建了BRD7的Bromodomain相关缺失突变体,通过肽段结合实验分析上述突变体与乙酰化组蛋白3结合的能力.结果表明,ZAloop与BCloop的完整性对于BRD7结合乙酰化的组蛋白3有着重要的意义.同时通过免疫荧光分析,证实了ZAloop与BCloop的完整性能够影响BRD7的亚细胞定位.最后,证实了BRD7与CBP可能存在交互作用.CBP不仅具有乙酰化转移酶活性(HATs),能够对组蛋白末端进行乙酰化修饰,并且作为一种重要的细胞转录因子广泛参与细胞的各种生物学活动.
Brominated region-containing protein 7 (BRD7) is a novel gene cloned using a cDNA-based differential analysis that has been shown to bind acetylated histone 3 with a recognition site at amino acid 14 of histone 3; and It was confirmed that the Bromodomain-deficient BRD7 mutant loses its ability to bind to acetylated histone 3. Bromodomain is an evolutionarily conserved functional domain that has distinct spatial conformations : Includes four left-handed, “helix (αz, αA, αB, αC)” and two “Zloop (BCloop)” antiparallel arrangement.By comprehensive analysis of bioinformatics, it is predicted that BRD7 may have the above According to the results of the above analysis, the Bromodomain-related deletion mutant of BRD7 was constructed and the ability of these mutants to bind to acetylated histone 3 was analyzed by peptide binding experiments. The results showed that the integrity of ZAloop and BCloop was not significantly affected by the acetylation of BRD7 Of histone 3 has an important significance.At the same time by immunofluorescence analysis showed that the integrity of ZAloop and BCloop can affect the BRD7 subcellular localization.Finally, it was confirmed that BRD7 and CBP may exist .CBP interaction not only acetyltransferase activity (HATs), capable of histone acetylation end, and as an important cellular transcription factor involved in a wide variety of biological activities of cells.