Regulation of myosin accumulation by muscle activity in cell culture.
Tetrodotoxin (TTX), at concentrations that do not interfere with normal myogenesis or with myosin synthesis, causes of cultured muscle fibres to accumulate myosin heavy chain peptides. This effect is now shown to be reversible. On removal of TTX, muscle fibres begin to reaccumulate myosin heavy chains and it appears that the myosin heavy chains display a 230% increase in stability when cells are shifted from TTX to a normal medium without TTX. Total protein stability or turnover is not affected by TTX. The ability of TTX to induce failure of accumulation of myosin heavy-chain in cultured muscle fibres does not extend to cultured chick fibroblasts. TTX also does not perturb normal uptake of [3H] leucine during a 1 h pulse and the leucine-specific activity within TTX-treated cells is essentially equivalent to that within normal cells. Finally, limited proteolysis of myosin heavy chain isolated from TTX-treated and normal muscle fibres and display of cleavage products on SDS-polyacrylamide gels does not reveal any significant difference between the two myosins. We conclude that failure of TTX muscle to accumulate myosin heavy chain is not related to impaired synthesis, to changes in myosin heavy-chain primary structure, or to overall changes in muscle fibre proteolytic activity. We speculate that the increase in degradation and resulting failure to accumulate myosin heavy chain in TTX cells is related to an inability of TTX-related muscle fibres to assemble newly synthesized fibrillar proteins into structures such as filaments or fibrils. Failure of assembly would lead to increased exposure to base-line levels of muscle proteolysis and to the observed lack of accumulation of myosin heavy chain.
D000818:Animals; D002478:Cells, Cultured; D002642:Chick Embryo; D004355:Drug Stability; D007700:Kinetics; D007930:Leucine / Q000378:metabolism; D009124:Muscle Proteins / Q000096:biosynthesis; D009132:Muscles / Q000187:drug effects / Q000196:embryology* / Q000378:metabolism; D009218:Myosins / Q000378:metabolism*; D013779:Tetrodotoxin / Q000494:pharmacology*
D009124:Muscle Proteins; D013779:Tetrodotoxin; D009218:Myosins; D007930:Leucine