Enhanced synthesis of a specific protein by 12-O-tetradecanoylphorbol-13-acetate in cultured chick embryo muscle cells.

The tumor promoter 12-O-tetradecanoylphorbol-13-acetate (TPA) induces in cultured postmitotic myotubes specific alterations of synthesized proteins as revealed by one-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis after pulse labeling with [35S]methionine. Synthesis of myosin heavy chain is remarkably inhibited after exposure to 1.6 X 10(-7) M TPA for periods of 9 hr or longer. During shorter periods of TPA treatment (2 hr), an enhanced synthesis of a Mr 31,000 polypeptide is observed, which is associated with the particulate fraction of cultured myotubes. "Pulse chase" experiments show that this polypeptide is not a degradation product induced by TPA. The stimulation of Mr 31,000 polypeptide requires simultaneous RNA synthesis, since actinomycin D completely and selectively abolishes [35S]methionine incorporation into this polypeptide. The stimulation of Mr 31,000 polypeptide is a transient biosynthetic event not detectable after prolonged incubation (24 hr) of myotubes with the tumor promoter. However, TPA-containing medium preincubated with cultures for up to 24 hr induces stimulation of Mr 31,000 polypeptide when administered to untreated cultures. The early stimulatory effect on Mr 31,000 polypeptide synthesis and the late inhibitory effects on contractile protein synthesis are also observed when postmitotic, unfused myoblasts, rather than myotubes, are treated with TPA.