title: TGF-beta receptor-mediated signalling through Smad2, Smad3 and Smad4.
abstract: Smad family members are newly identified essential intracellular signalling components of the transforming growth factor-beta (TGF-beta) superfamily. Smad2 and Smad3 are structurally highly similar and mediate TGF-beta signals. Smad4 is distantly related to Smads 2 and 3, and forms a heteromeric complex with Smad2 after TGF-beta or activin stimulation. Here we show that Smad2 and Smad3 interacted with the kinase-deficient TGF-beta type I receptor (TbetaR)-I after it was phosphorylated by TbetaR-II kinase. TGF-beta1 induced phosphorylation of Smad2 and Smad3 in Mv1Lu mink lung epithelial cells. Smad4 was found to be constitutively phosphorylated in Mv1Lu cells, the phosphorylation level remaining unchanged upon TGF-beta1 stimulation. Similar results were obtained using HSC4 cells, which are also growth-inhibited by TGF-beta. Smads 2 and 3 interacted with Smad4 after TbetaR activation in transfected COS cells. In addition, we observed TbetaR-activation-dependent interaction between Smad2 and Smad3. Smads 2, 3 and 4 accumulated in the nucleus upon TGF-beta1 treatment in Mv1Lu cells, and showed a synergistic effect in a transcriptional reporter assay using the TGF-beta-inducible plasminogen activator inhibitor-1 promoter. Dominant-negative Smad3 inhibited the transcriptional synergistic response by Smad2 and Smad4. These data suggest that TGF-beta induces heteromeric complexes of Smads 2, 3 and 4, and their concomitant translocation to the nucleus, which is required for efficient TGF-beta signal transduction.
authors.affiliation: Ludwig Institute for Cancer Research, Box 595, S-751 24 Uppsala, Sweden.
authors.forename: A
authors.initials: A
authors.lastname: Nakao
authors.forename: T
authors.initials: T
authors.lastname: Imamura
authors.forename: S
authors.initials: S
authors.lastname: Souchelnytskyi
authors.forename: M
authors.initials: M
authors.lastname: Kawabata
authors.forename: A
authors.initials: A
authors.lastname: Ishisaki
authors.forename: E
authors.initials: E
authors.lastname: Oeda
authors.forename: K
authors.initials: K
authors.lastname: Tamaki
authors.forename: J
authors.initials: J
authors.lastname: Hanai
authors.forename: C H
authors.initials: CH
authors.lastname: Heldin
authors.forename: K
authors.initials: K
authors.lastname: Miyazono
authors.forename: P
authors.initials: P
authors.lastname: ten Dijke
mesh_terms: D030201:Activin Receptors, Type I; D000595:Amino Acid Sequence; D000818:Animals; D000918:Antibody Specificity; D001692:Biological Transport; D019556:COS Cells; D002467:Cell Nucleus / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000276:immunology / Q000378:metabolism*; D004847:Epithelial Cells; D017930:Genes, Reporter; D006801:Humans; D008168:Lung / Q000166:cytology; D008907:Mink; D008954:Models, Biological; D008969:Molecular Sequence Data; D010766:Phosphorylation; D011485:Protein Binding; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism; D000077293:Receptor, Transforming Growth Factor-beta Type I; D000077294:Receptor, Transforming Growth Factor-beta Type II; D018125:Receptors, Transforming Growth Factor beta / Q000378:metabolism*; D015398:Signal Transduction; D051899:Smad2 Protein; D051900:Smad3 Protein; D051901:Smad4 Protein; D015534:Trans-Activators / Q000276:immunology / Q000378:metabolism*; D014158:Transcription, Genetic; D014407:Tumor Cells, Cultured
chemical_list: D004268:DNA-Binding Proteins; D018125:Receptors, Transforming Growth Factor beta; C497782:SMAD2 protein, human; C497790:SMAD3 protein, human; C497796:SMAD4 protein, human; D051899:Smad2 Protein; D051900:Smad3 Protein; D051901:Smad4 Protein; D015534:Trans-Activators; D017346:Protein Serine-Threonine Kinases; D030201:Activin Receptors, Type I; D000077293:Receptor, Transforming Growth Factor-beta Type I; D000077294:Receptor, Transforming Growth Factor-beta Type II