Transforming growth factor (TGF)- is a multifunctional growth factor with important roles in development, cell proliferation, and matrix deposition. It signals through the sequential activation of two serine/ threonine kinase receptors, the type I and type II receptors. A third cell surface receptor, betaglycan, serves as a co-receptor for TGF- in some cell types, enhancing TGF--mediated signaling. We have examined the function of betaglycan in renal epithelial LLC-PK 1 cells that lack endogenous betaglycan. We demonstrate that the expression of betaglycan in LLC-PK 1 cells results in inhibition of TGF- signaling as measured by reporter gene expression, thymidine incorporation, collagen production, and phosphorylation of the downstream signaling effectors Smad2 and Smad3. In comparison, the expression of betaglycan in L6 myoblasts enhances TGF- signaling, which is consistent with the published literature. The effects of betaglycan in LLC-PK 1 cells are not mediated by ligand sequestration or increased production of a soluble form of the receptor, which has been reported to serve as a ligand antagonist. We demonstrate instead that in LLC-PK 1 cells, unlike L6 cells, expression of betaglycan prevents association between the type I and type II TGF- receptors, which is required for signaling. This is a function of the glycosaminoglycan modifications of betaglycan. Betaglycan in LLC-PK 1 cells exhibits higher molecular weight glycosaminoglycan (GAG) chains than in L6 cells, and a GAG ؊ betaglycan mutant does not inhibit TGF- signaling or type I/type II receptor association in LLC-PK 1 cells. Our data indicate that betaglycan can function as a potent inhibitor of TGF- signaling by a novel mechanism and provide support for an essential but complex role for proteoglycan co-receptors in growth factor signaling.
TGF-1 is an essential regulator of development, cell proliferation, and matrix deposition (1-3). Biological effects of TGF- are achieved through the sequential activation of two high affinity serine/threonine kinase receptors, the type II (TRII) and the type I (TRI) TGF- receptors. TGF- signaling is initiated by the binding of TGF- to TRII followed by the formation or stabilization of TRI-TRII complexes, transphosphorylation of TRI by TRII, and phosphorylation of the receptor-associated cytoplasmic effector molecules Smad2 and Smad3 by TRI (1, 3). A third TGF- transmembrane receptor, the type III receptor or betaglycan, is a widely expressed heparan and chondroitin sulfate proteoglycan that binds TGF- with high affinity through its protein core but has no apparent intrinsic signaling activity (4 -7).Betaglycan is believed to be a co-receptor for TGF-. In several cell lines, including rat L6 myoblasts lacking endogenous betaglycan, it significantly increases the affinity of TRI and TRII for TGF-, thereby enhancing the response to TGF- (4, 5). In addition, betaglycan eliminates differences in efficacy among the three TGF- isoforms. TGF-2, which binds to TRI and TRII with low affinity ...