Cytochalasin D alters the rate of synthesis of some HEp‐2 cytoskeletal proteins

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Abstract
1. 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. 2. After treatment of HEp-2 cells with 0.2 .mu.M or 2.0 .mu.M 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.