THE GLI GENE ENCODES A NUCLEAR-PROTEIN WHICH BINDS SPECIFIC SEQUENCES IN THE HUMAN GENOME
THE GLI GENE ENCODES A NUCLEAR-PROTEIN WHICH BINDS SPECIFIC SEQUENCES IN THE HUMAN GENOME
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DOI：
10.1128/mcb.10.2.634
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发表时间：
1990-02-01
影响因子：
5.3
通讯作者：
VOGELSTEIN, B
中科院分区：
文献类型：
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作者：
KINZLER, KW;VOGELSTEIN, B
GLI基因在人类肿瘤的一个子集中扩增，并编码具有五个锌指DNA结合基序的蛋白质产物。在这项研究中，我们表明，GLI基因产物具有主要的核定位，并结合DNA的序列特异性的方式。三个GLI结合位点被确定通过使用一种新的程序，其中总的人DNA结合到GLI重组融合蛋白，和聚合酶链反应被用来扩增和恢复结合的序列。GLI蛋白保护所有三个结合位点内的23至24个碱基的区域，并且在每种情况下保护的区域包括9个碱基对的序列5“-GACCACCCA-3”。其中一个结合位点包含在可变数目的串联重复类型的63个碱基对重复内，而另外两个位点在基因组中仅出现一次。本文所用的GLI结合位点的鉴定方法也适用于其它锌指蛋白的鉴定。
The GLI gene is amplified in a subset of human tumors and encodes a protein product with five zinc finger DNA-binding motifs. In this study, we show that the GLI gene product has a predominantly nuclear localization and binds dNA in a sequence-specific fashion. Three GLI binding sites were identified by using a novel procedure in which total human DNA was bound to a GLI recombinant fusion protein, and the polymerase chain reaction was used to amplify and recover the bound sequences. The GLI protein protected a 23- to 24-base region within all three binding sites, and the protected region in each case included the 9-base-pair sequences 5''-GACCACCCA-3''. One of the binding sites was contained within a 63-base-pair repeat of the variable number of tandem repeat type, whereas the other two sites were represented once in the genome. The approach used here to identify GLI binding sites should be applicable to the characterization of othe zinc finger proteins.