Transforming growth factor beta1 signaling via interaction with cell surface Hyal-2 and recruitment of WWOX/WOX1.
Transforming growth factor beta (TGF-beta) initiates multiple signal pathways and activates many downstream kinases. Here, we determined that TGF-beta1 bound cell surface hyaluronidase Hyal-2 on microvilli in type II TGF-beta receptor-deficient HCT116 cells, as determined by immunoelectron microscopy. This binding resulted in recruitment of proapoptotic WOX1 (also named WWOX or FOR) and formation of Hyal-2.WOX1 complexes for relocation to the nuclei. TGF-beta1 strengthened the binding of the catalytic domain of Hyal-2 with the N-terminal Tyr-33-phosphorylated WW domain of WOX1, as determined by time lapse fluorescence resonance energy transfer analysis in live cells, co-immunoprecipitation, and yeast two-hybrid domain/domain mapping. In promoter activation assay, ectopic WOX1 or Hyal-2 alone increased the promoter activity driven by Smad. In combination, WOX1 and Hyal-2 dramatically enhanced the promoter activation (8-9-fold increases), which subsequently led to cell death (>95% of promoter-activated cells). TGF-beta1 supports L929 fibroblast growth. In contrast, transiently overexpressed WOX1 and Hyal-2 sensitized L929 to TGF-beta1-induced apoptosis. Together, TGF-beta1 invokes a novel signaling by engaging cell surface Hyal-2 and recruiting WOX1 for regulating the activation of Smad-driven promoter, thereby controlling cell growth and death.
Hsu
Li-Jin
LJ
Institute of Molecular Medicine, National Cheng Kung University Medical College, Tainan 70101, Taiwan. wox1world@gmail.com
Schultz
Lori
L
Hong
Qunying
Q
Van Moer
Kris
K
Heath
John
J
Li
Meng-Yen
MY
Lai
Feng-Jie
FJ
Lin
Sing-Ru
SR
Lee
Ming-Hui
MH
Lo
Cheng-Peng
CP
Lin
Yee-Shin
YS
Chen
Shur-Tzu
ST
Chang
Nan-Shan
NS
D000818:Animals; D017209:Apoptosis; D015815:Cell Adhesion Molecules / Q000235:genetics / Q000502:physiology*; D002462:Cell Membrane / Q000502:physiology*; D015179:Colorectal Neoplasms; D017931:DNA Primers; D031541:Fluorescence Resonance Energy Transfer; D058851:GPI-Linked Proteins; D005786:Gene Expression Regulation; D045325:HCT116 Cells; D006801:Humans; D006821:Hyaluronoglucosaminidase / Q000235:genetics / Q000502:physiology*; D007739:L Cells; D051379:Mice; D010088:Oxidoreductases / Q000502:physiology*; D011401:Promoter Regions, Genetic; D017346:Protein Serine-Threonine Kinases / Q000172:deficiency / Q000502:physiology*; D051381:Rats; D000077293:Receptor, Transforming Growth Factor-beta Type I; D000077294:Receptor, Transforming Growth Factor-beta Type II; D018125:Receptors, Transforming Growth Factor beta / Q000172:deficiency / Q000502:physiology*; D025521:Tumor Suppressor Proteins / Q000502:physiology*; D000074584:WW Domain-Containing Oxidoreductase
D015815:Cell Adhesion Molecules; D017931:DNA Primers; D058851:GPI-Linked Proteins; D018125:Receptors, Transforming Growth Factor beta; D025521:Tumor Suppressor Proteins; D010088:Oxidoreductases; D000074584:WW Domain-Containing Oxidoreductase; C486071:WWOX protein, human; D017346:Protein Serine-Threonine Kinases; D000077293:Receptor, Transforming Growth Factor-beta Type I; D000077294:Receptor, Transforming Growth Factor-beta Type II; C000625843:Tgfbr1 protein, rat; C473096:Hyal2 protein, human; D006821:Hyaluronoglucosaminidase