Adenovirus-mediated transfer of a truncated transforming growth factor-beta (TGF-beta) type II receptor completely and specifically abolishes diverse signaling by TGF-beta in vascular wall cells in primary culture.
We constructed an adenoviral vector expressing a mutated human type II transforming growth factor-beta (TGF-beta) receptor that was truncated of its kinase domain (AdexCATbetaTR) and examined whether this truncated receptor could abolish signaling by TGF-beta using arterial endothelial cells and smooth muscle cells, as well as a lung epithelial cell line (Mv1Lu). Infection of cells with AdexCATbetaTR induced expression of the truncated receptor, the amount of which would be excessive compared with those of both full-length type I and type II receptors, as assessed by levels of their mRNAs. The antiproliferative effect of TGF-beta was completely eliminated in both endothelial cells and Mv1Lu that were infected with AdexCATbetaTR. The transcriptional activation by TGF-beta of plasminogen activator inhibitor-1 and fibronectin was entirely suppressed. Abrogation of the TGF-beta-enhanced production of type I collagen in infected smooth muscle cells was confirmed by immunocytostaining and by [14C]proline incorporation in a quantitative manner. Mitogenic response to other growth factors remained unaffected in infected cells. Our data demonstrated that the adenovirus-mediated transfer of a truncated type II TGF-beta receptor completely and specifically abolishes the diverse effects of TGF-beta as a dominant-negative mutation, supporting the hypothesis that both the type I and type II receptors are required for all signaling by TGF-beta. This method may facilitate the clarification of the role of TGF-beta both in vitro and in vivo.
Yamamoto
H
H
Molecular Cardiology Unit, Research Institute of Angiocardiology, Kyushu University School of Medicine, Fukuoka, 812-82, Japan.
Ueno
H
H
Ooshima
A
A
Takeshita
A
A
D000256:Adenoviridae; D000818:Animals; D001011:Aorta; D001013:Aorta, Thoracic; D015152:Blotting, Northern; D002417:Cattle; D002455:Cell Division / Q000187:drug effects; D002460:Cell Line; D002478:Cells, Cultured; D004730:Endothelium, Vascular / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D005353:Fibronectins / Q000096:biosynthesis; D005822:Genetic Vectors; D006801:Humans; D008168:Lung; D008907:Mink; D009131:Muscle, Smooth, Vascular / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D017395:Plasminogen Activator Inhibitor 1 / Q000096:biosynthesis; D017346:Protein Serine-Threonine Kinases; D011505:Protein-Tyrosine Kinases / Q000096:biosynthesis / Q000235:genetics; D000077294:Receptor, Transforming Growth Factor-beta Type II; D018125:Receptors, Transforming Growth Factor beta / Q000096:biosynthesis / Q000187:drug effects / Q000502:physiology*; D011994:Recombinant Proteins / Q000187:drug effects / Q000378:metabolism; D017384:Sequence Deletion; D015398:Signal Transduction; D014158:Transcription, Genetic; D015533:Transcriptional Activation / Q000187:drug effects; D014162:Transfection; D016212:Transforming Growth Factor beta / Q000494:pharmacology*
D005353:Fibronectins; D017395:Plasminogen Activator Inhibitor 1; D018125:Receptors, Transforming Growth Factor beta; D011994:Recombinant Proteins; D016212:Transforming Growth Factor beta; D011505:Protein-Tyrosine Kinases; D017346:Protein Serine-Threonine Kinases; D000077294:Receptor, Transforming Growth Factor-beta Type II