title: Mode and specificity of binding of the small molecule GANT61 to GLI determines inhibition of GLI-DNA binding.
abstract: 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.
authors: Department of Cancer Biology, Lerner Research Institute, Cleveland Clinic, Cleveland, OH.
authors: Akwasi
authors: A
authors: Agyeman
authors: Babal K
authors: BK
authors: Jha
authors: Tapati
authors: T
authors: Mazumdar
authors: Department of Cancer Biology, Lerner Research Institute, Cleveland Clinic, Cleveland, OH.
authors: Janet A
authors: JA
authors: Houghton
mesh_terms: D045744:Cell Line, Tumor; D002470:Cell Survival; D003110:Colonic Neoplasms / Q000378:metabolism*; D053823:Hedgehog Proteins / Q000378:metabolism*; D006801:Humans; D000090062:Kruppel-Like Factor 4; D011725:Pyridines / Q000378:metabolism*; D011743:Pyrimidines / Q000378:metabolism*; D015398:Signal Transduction; D014157:Transcription Factors / Q000235:genetics* / Q000378:metabolism*; D014162:Transfection; D000071676:Zinc Finger Protein GLI1
chemical_list: C551027:GANT 61; C400551:GLI1 protein, human; D053823:Hedgehog Proteins; C000715467:KLF4 protein, human; D000090062:Kruppel-Like Factor 4; D011725:Pyridines; D011743:Pyrimidines; D014157:Transcription Factors; D000071676:Zinc Finger Protein GLI1