Mode and specificity of binding of the small molecule GANT61 to GLI determines inhibition of GLI-DNA binding.
Mode and specificity of binding of the small molecule GANT61 to GLI determines inhibition of GLI-DNA binding.
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DOI：
10.18632/oncotarget.2046
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发表时间：
2014-06-30
期刊：
影响因子：
--
通讯作者：
Houghton JA
中科院分区：
文献类型：
--
作者：
Agyeman A;Jha BK;Mazumdar T;Houghton JA
GLI基因GLI 1和GLI 2是调节经典Hedgehog（HH）信号传导途径（SHH->PTCH->SMO->GLI）远端靶基因的转录因子，在胚胎发育、组织图案化和分化中受到严格调节。GLI 1和GLI 2都是癌基因，在许多类型的人类癌症中组成性激活。在结肠癌细胞中，致癌KRAS-GLI信号传导绕过HH-SMO-GLI轴以在靶基因的转录调节中引导并激活GLI。我们使用小分子GLI抑制剂GANT 61在一组7种人结肠癌细胞系中观察到广泛的细胞死亡。通过计算对接和表面等离子体共振的实验确认，GANT 61在位点E119和E167处与锌指2和3之间的5-锌指GLI 1蛋白结合，独立于GLI-DNA结合区域，并且在GLI 1和GLI 2之间保守。GANT 61不与其他锌指转录因子（KLF 4、TFIIβ）结合。突变GLI 1中预测的GANT 61结合位点显著抑制GANT 61-GLI结合和GLI-荧光素酶活性。数据确立了GANT 61靶向GLI的特异性，并证实了GLI在癌细胞存活中的关键作用。因此，在癌症治疗中靶向GLI可能具有高影响。
The GLI genes, GLI1 and GLI2, are transcription factors that regulate target genes at the distal end of the canonical Hedgehog (HH) signaling pathway (SHH->PTCH->SMO->GLI), tightly regulated in embryonic development, tissue patterning and differentiation. Both GLI1 and GLI2 are oncogenes, constitutively activated in many types of human cancers. In colon cancer cells oncogenic KRAS-GLI signaling circumvents the HH-SMO-GLI axis to channel through and activate GLI in the transcriptional regulation of target genes. We have observed extensive cell death in a panel of 7 human colon carcinoma cell lines using the small molecule GLI inhibitor GANT61. Using computational docking and experimental confirmation by Surface Plasmon Resonance, GANT61 binds to the 5-zinc finger GLI1 protein between zinc fingers 2 and 3 at sites E119 and E167, independent of the GLI-DNA binding region, and conserved between GLI1 and GLI2. GANT61 does not bind to other zinc finger transcription factors (KLF4, TFIIβ). Mutating the predicted GANT61 binding sites in GLI1 significantly inhibits GANT61-GLI binding and GLI-luciferase activity. Data establish the specificity of GANT61 for targeting GLI, and substantiate the critical role of GLI in cancer cell survival. Thus, targeting GLI in cancer therapeutics may be of high impact.
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影响因子：
3.7
作者：
Hofmann I;Weiss A;Elain G;Schwaederle M;Sterker D;Romanet V;Schmelzle T;Lai A;Brachmann SM;Bentires-Alj M;Roberts TM;Sellers WR;Hofmann F;Maira SM
通讯作者：
Maira SM
影响因子：
64.8
作者：
KINZLER, KW;RUPPERT, JM;VOGELSTEIN, B
通讯作者：
VOGELSTEIN, B
影响因子：
8
作者：
Day, YSN;Baird, CL;Myszka, DG
通讯作者：
Myszka, DG
影响因子：
4.8
作者：
Ji, Zhenyu;Mei, Fang C.;Cheng, Xiaodong
通讯作者：
Cheng, Xiaodong
影响因子：
4.3
作者：
Lauth, Matthias;Toftgard, Rune
通讯作者：
Toftgard, Rune