Differential effects of the Ski-interacting protein (SKIP) on differentiation induced by transforming growth factor-beta1 and bone morphogenetic protein-2 in C2C12 cells.

The transforming growth factor-beta (TGF-beta) and bone morphogenetic proteins (BMP) are key regulatory factors that affect many critical cellular events in growth and development. Recently, we have shown that the Ski-interacting protein (SKIP) can augment TGF-beta signals. Here, we extended these studies by examining the biologic consequences of SKIP overexpression on TGF-beta1 and BMP-2 signals in C2C12 cells. C2C12 myoblasts differentiate into myotubes when the media is depleted of mitogenic factors, and TGF-beta1 inhibits this myotube formation. BMP-2 not only inhibits the myotube formation, but also induces C2C12 cells to differentiate into osteoblasts. Here, we show that SKIP-overexpressing C2C12 cells treated with TGF-beta1 or BMP-2 displayed no differences in comparison to vector control cells in their ability to form myotubes or in the expression of the myogenic markers myosin heavy chain-1 and myogenin. Unexpectedly, SKIP-overexpressing C2C12 cells treated with BMP-2 displayed suppressed expression of the induced osteoblast markers alkaline phosphatase, osteocalcin, and the transcription factor Runx2. Lastly, SKIP could repress transcription induced by BMP-2 in luciferase reporter assays done in C2C12 cells. These data show that SKIP has specific inhibitory effects on BMP-2-induced differentiation and implicate SKIP to be a novel regulator of the differentiation programming induced by TGF-beta signals.