title: Hyaluronan activates Hyal-2/WWOX/Smad4 signaling and causes bubbling cell death when the signaling complex is overexpressed.
abstract: Malignant cancer cells frequently secrete significant amounts of transforming growth factor beta (TGF-β), hyaluronan (HA) and hyaluronidases to facilitate metastasizing to target organs. In a non-canonical signaling, TGF-β binds membrane hyaluronidase Hyal-2 for recruiting tumor suppressors WWOX and Smad4, and the resulting Hyal-2/WWOX/Smad4 complex is accumulated in the nucleus to enhance SMAD-promoter dependent transcriptional activity. Yeast two-hybrid analysis showed that WWOX acts as a bridge to bind both Hyal-2 and Smad4. When WWOX-expressing cells were stimulated with high molecular weight HA, an increased formation of endogenous Hyal-2/WWOX/Smad4 complex occurred rapidly, followed by relocating to the nuclei in 20-40 min. In WWOX-deficient cells, HA failed to induce Smad2/3/4 relocation to the nucleus. To prove the signaling event, we designed a real time tri-molecular FRET analysis and revealed that HA induces the signaling pathway from ectopic Smad4 to WWOX and finally to p53, as well as from Smad4 to Hyal-2 and then to WWOX. An increased binding of the Smad4/Hyal-2/WWOX complex occurs with time in the nucleus that leads to bubbling cell death. In contrast, HA increases the binding of Smad4/WWOX/p53, which causes membrane blebbing but without cell death. In traumatic brain injury-induced neuronal death, the Hyal-2/WWOX complex was accumulated in the apoptotic nuclei of neurons in the rat brains in 24 hr post injury, as determined by immunoelectron microscopy. Together, HA activates the Hyal-2/WWOX/Smad4 signaling and causes bubbling cell death when the signaling complex is overexpressed.
authors.affiliation: Guthrie Research Institute, Laboratory of Molecular Immunology, Sayre, PA, USA.|Department of Medical Laboratory Science and Biotechnology, National Cheng Kung University College of Medicine, Tainan, Taiwan, ROC.
authors.forename: Li-Jin
authors.initials: LJ
authors.lastname: Hsu
authors.affiliation: Guthrie Research Institute, Laboratory of Molecular Immunology, Sayre, PA, USA.
authors.forename: Qunying
authors.initials: Q
authors.lastname: Hong
authors.affiliation: Guthrie Research Institute, Laboratory of Molecular Immunology, Sayre, PA, USA.|Department of Cell Biology and Anatomy, National Cheng Kung University College of Medicine, Tainan, Taiwan, ROC.
authors.forename: Shur-Tzu
authors.initials: ST
authors.lastname: Chen
authors.affiliation: Institute of Molecular Medicine, National Cheng Kung University College of Medicine, Tainan, Taiwan, ROC.
authors.forename: Hsiang-Lin
authors.initials: HL
authors.lastname: Kuo
authors.affiliation: Guthrie Research Institute, Laboratory of Molecular Immunology, Sayre, PA, USA.
authors.forename: Lori
authors.initials: L
authors.lastname: Schultz
authors.affiliation: Guthrie Research Institute, Laboratory of Molecular Immunology, Sayre, PA, USA.
authors.forename: John
authors.initials: J
authors.lastname: Heath
authors.affiliation: Institute of Molecular Medicine, National Cheng Kung University College of Medicine, Tainan, Taiwan, ROC.
authors.forename: Sing-Ru
authors.initials: SR
authors.lastname: Lin
authors.affiliation: Institute of Molecular Medicine, National Cheng Kung University College of Medicine, Tainan, Taiwan, ROC.
authors.forename: Ming-Hui
authors.initials: MH
authors.lastname: Lee
authors.affiliation: Institute of Molecular Medicine, National Cheng Kung University College of Medicine, Tainan, Taiwan, ROC.
authors.forename: Dong-Zhang
authors.initials: DZ
authors.lastname: Li
authors.affiliation: Institute of Molecular Medicine, National Cheng Kung University College of Medicine, Tainan, Taiwan, ROC.
authors.forename: Zih-Ling
authors.initials: ZL
authors.lastname: Li
authors.affiliation: Department of Medical Laboratory Science and Biotechnology, National Cheng Kung University College of Medicine, Tainan, Taiwan, ROC.
authors.forename: Hui-Ching
authors.initials: HC
authors.lastname: Cheng
authors.affiliation: Glycomed Research Inc., Hastings on Hudson, New York, NY, USA.
authors.forename: Gerard
authors.initials: G
authors.lastname: Armand
authors.affiliation: Guthrie Research Institute, Laboratory of Molecular Immunology, Sayre, PA, USA.|Institute of Molecular Medicine, National Cheng Kung University College of Medicine, Tainan, Taiwan, ROC.|Advanced Optoelectronic Technology Center, National Cheng Kung University, Tainan, Taiwan, ROC.|Graduate Institute of Biomedical Sciences, College of Medicine, China Medical University, Taichung, Taiwan, ROC.
authors.forename: Nan-Shan
authors.initials: NS
authors.lastname: Chang
mesh_terms: D000818:Animals; D016923:Cell Death / Q000502:physiology*; D045744:Cell Line, Tumor; D004195:Disease Models, Animal; D031541:Fluorescence Resonance Energy Transfer; D006801:Humans; D006820:Hyaluronic Acid / Q000378:metabolism*; D047468:Immunoprecipitation; D008856:Microscopy, Fluorescence; D016253:Microscopy, Immunoelectron; D009369:Neoplasms / Q000378:metabolism / Q000473:pathology*; D010088:Oxidoreductases / Q000378:metabolism*; D051381:Rats; D015398:Signal Transduction / Q000502:physiology*; D051901:Smad4 Protein / Q000378:metabolism*; D025521:Tumor Suppressor Proteins / Q000378:metabolism*; D020798:Two-Hybrid System Techniques; D000074584:WW Domain-Containing Oxidoreductase
chemical_list: D051901:Smad4 Protein; D025521:Tumor Suppressor Proteins; D006820:Hyaluronic Acid; D010088:Oxidoreductases; D000074584:WW Domain-Containing Oxidoreductase; C486071:WWOX protein, human
keywords: Hyal-2; Smad; WWOX; hyaluronan; hyaluronidase