Functional analysis of a novel KRAB/C2H2 zinc finger protein Mipu1
Functional analysis of a novel KRAB/C2H2 zinc finger protein Mipu1
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新型 KRAB/C2H2 锌指蛋白 Mipu1 的功能分析。
DOI：
10.1016/j.bbrc.2007.02.138
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发表时间：
2007-05-18
影响因子：
3.1
通讯作者：
Xiao, Xianzhong
中科院分区：
文献类型：
--
作者：
Jiang, Lei;Tang, Daolin;Xiao, Xianzhong
一个新的大鼠基因Mipu 1编码一个608个氨基酸的蛋白质，具有氨基端KRAB结构域和14个羧基端C2 H2锌指基序。Mipu 1通过其KRAB结构域或邻近其锌指区的接头定位于细胞核。利用与谷胱甘肽-琼脂糖凝胶珠结合的GST-Mipu 1，从随机寡核苷酸文库中提取共有的推定DNA结合位点（5 '-TGTCTTATCGAA-3'）。EMSA和靶点检测实验表明，含有该位点的探针能与纯化的GST-Mipu 1融合蛋白结合。将含有推定位点的寡核苷酸插入pGL 3启动子载体中以产生报告构建体。Mipu 1的过表达抑制了报告基因的表达，并呈剂量依赖性。共有序列的突变分析表明，Mipu 1介导的抑制是序列依赖性的。这些结果表明，Mipu 1是一个核蛋白，其功能作为一个转录抑制因子。(c)2007爱思唯尔公司All rights reserved.
A novel rat gene, Mipu1, encodes a 608 amino acid protein with an amino- terminal KRAB domain and 14 carboxyl-terminal C2H2 zinc finger motifs. Mipu1 is localized to the nucleus through its KRAB domain or the linker adjacent to its zinc finger region. Using the GST-Mipu1 bound to glutathione-Sepharose beads, a consensus putative DNA binding site (5'-TGTCTTATCGAA-3') was extracted from a random oligonucleotide library. EMSA and target detection assay showed that the probe containing the putative site can bind to purified GST-Mipu1 fusion protein. The oligonucleotide containing the putative site was inserted into the pGL3-promotor vector to produce a reporter construct. The expression of reporter gene was repressed by overexpression of Mipu1 in a dose-dependent manner. Mutation analysis of the consensus sequence indicated that the repression mediated by Mipu1 is sequence-dependent. These results suggest that Mipu1 is a nuclear protein, which functions as a transcriptional repressor. (c) 2007 Elsevier Inc. All rights reserved.