Messenger RNA for a phorbol-ester induced 48,000 dalton protein from human melanoma cells.
Messenger RNA of the phorbol ester-induced 48kDa protein from human melanoma cells (Bowes) was isolated, characterized and used to study the protein processing. The 48kDa mRNA is induced simultaneously with that of tissue-type plasminogen activator. This induction is prominent as shown by sedimentation profiles on linear sucrose gradients. The mRNA can be isolated by classical phenol extractions, has a poly(A)-tail and sediments with a coefficient of 20 S. Translation in reticulocyte lysates yields a 48kDa protein whether the translation is modified with canine pancreas microsomal membranes or not. Analysis of 48kDa mRNA translation products by sodium dodecyl sulphate/polyacrylamide gel electrophoresis showed that the phorbol ester-induced 48kDa is a monomeric one-chain polypeptide. Glycosylation could not be detected, nor signal peptide cleaving, suggesting that it is a non-secreted intracellular protein.
D000818:Animals; D001345:Autoradiography; D002460:Cell Line; D002499:Centrifugation, Density Gradient; D006801:Humans; D008545:Melanoma / Q000235:genetics*; D008970:Molecular Weight; D009865:Oocytes / Q000378:metabolism; D010703:Phorbol Esters / Q000494:pharmacology*; D014176:Protein Biosynthesis / Q000187:drug effects; D012333:RNA, Messenger / Q000378:metabolism*; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D013997:Time Factors; D010959:Tissue Plasminogen Activator / Q000032:analysis; D014981:Xenopus
D010703:Phorbol Esters; D012333:RNA, Messenger; D010959:Tissue Plasminogen Activator; D013755:Tetradecanoylphorbol Acetate