Cytochalasin D alters the rate of synthesis of some HEp-2 cytoskeletal proteins. Examination by two-dimensional gel electrophoresis.
The most abundant proteins of HEp-2 cells were resolved by two-dimensional gel electrophoresis. The protein spots corresponding to several cytoskeletal proteins (vimentin, alpha-tubulin, beta-tubulin, alpha-actinin, tropomyosins, and cytokeratins) were identified by comigration with protein markers or by immunological techniques. After treatment of HEp-2 cells with 0.2 microM or 2.0 microM cytochalasin D for 20 h, radioautograms of two-dimensional gel patterns of lysates from cells pulse-labeled with [35S]methionine indicated that the drug altered the rate of synthesis of some proteins. The relative rate of synthesis of the identified cytoskeletal proteins was measured. Synthesis of alpha-actinin, the higher-molecular-mass pair of tropomyosins and actin were similarly increased with cytochalasin D treatment, suggesting coordinate induction. Vimentin and tubulin synthesis was depressed. One cytokeratin exhibited an increase in synthesis comparable to actin, another was increased to a lesser extent and one was decreased.
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Tannenbaum
D000185:Actinin / Q000096:biosynthesis; D001345:Autoradiography; D002460:Cell Line; D015638:Cytochalasin D; D003572:Cytochalasins / Q000494:pharmacology*; D003598:Cytoskeletal Proteins / Q000096:biosynthesis*; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D007120:Immunochemistry; D007633:Keratins / Q000096:biosynthesis; D010452:Peptide Biosynthesis; D014335:Tropomyosin / Q000096:biosynthesis; D014404:Tubulin / Q000096:biosynthesis; D014746:Vimentin / Q000096:biosynthesis
D003572:Cytochalasins; D003598:Cytoskeletal Proteins; D014335:Tropomyosin; D014404:Tubulin; D014746:Vimentin; D000185:Actinin; D015638:Cytochalasin D; D007633:Keratins