TGF-α-induced breakdown of stree fibers and degradation of tropmyosin in NRK cells is blocked by a proteasome inhibitor

Robert H. Warren

Research output: Contribution to journalArticle

12 Scopus citations

Abstract

Treatment of NRK cells with TGF-α in the presence of serum initiates disassembly of cytoskeletal stress fibers and suppresses the synthesis of tropomyosin isoforms (TMs) 1, 2, and 3 but not TMs 4 and 5 (Copper et al., Cancer Res. 47, 4493-4500, 1987). In order to determine how the loss of tropomyosin is induced and what role it plays in cytoskeletal disruption, the turnover of tropomyosin was studied in the presence of the transforming growth factor and protease inhibitors. Cells were pulse-labeled with [35S]methionine and chased in the absence or the presence of the growth factor. It was found that TMs 1, 2, and 3 are degraded at about twice the rate of TMs 4 and 5 in control cells and that the rate of degradation of TMs 1-3 is accelerated by the growth factors. Degradation of TMs in control and growth factor-treated cells is blocked by a membrane-permeable inhibitor of cysteine proteases (LLnL) that acts upon calpains and proteasomes, and the cells maintain a flattened shape with a normal complement of stress fibers. Application of inhibitors that block calpains but not proteasomes does not block TM degradation. Treatments (suspension culture or cytochalasin B) that disrupt stress fibers without application of the growth factors also accelerate TM degradation, suggesting that acceleration of TM degradation is a consequence of its release from stress fibers during their breakdown. The normally more rapid turnover of the TM isoforms 1-3 that are lost in the phenotypically transformed cells could serve to facilitate the cytoskeletal reorganization that follows the activition of signal transduction pathways by the transforming growth factors observed in this study or during other rearrangements of the cytoskeleton such as occur during cell migration or mitosis.

Original languageEnglish (US)
Pages (from-to)294-303
Number of pages10
JournalExperimental Cell Research
Volume236
Issue number1
DOIs
StatePublished - Oct 10 1997

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Keywords

  • Calpain
  • LLnL
  • Proteasome
  • Stress fibers
  • Transforming growth factor-α (TGF- α)
  • Tropomyosin

ASJC Scopus subject areas

  • Cell Biology

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