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. 1975 Dec 1;67(3):523–537. doi: 10.1083/jcb.67.3.523

Synthesis of myosin heavy and light chains in muscle cultures

PMCID: PMC2111676  PMID: 127801

Abstract

The weight ratio of myosin/actin, the myosin heavy chain content as the percentage of total protein (wt/wt), and the kinds of myosin light chains were determined in (a) standard muscle cultures, (b) pure myotube cultures, and (c) fibroblast cultures. Cells for these cultures were obtained from the breast of 11-day chick embryos. Standard cultures contain, in addition to myotubes, large numbers of replicating mononucleated cells. By killing these replicating cells with cytosine arabinoside, pure myotube cultures were obtained. The myosin/actin ratio (wt/wt) for pure myotube, standard muscle, and fibroblast cultures average 3.1, 1.9, and 1.1 respectively. By day 7, myosin in myotube cultures represents a minimum of 7% of the total protein, but about 3% in standard cultures and less than 1.5% in fibroblasts cultures. Myosin from standard cultures contains light chain LC1, LC2, and LC3, with a relative stoichiometry of the molarity of 1.0:1.9:0.5 and mol wt of 25,000, 18,000 and 16,000 daltons, identical to those in adult fast muscle. Myosin from pure myotubes exhibits light chains LC1 and LC2, with a molar ratio of 1.5:1.6. Myosin from fibroblast cultures possesses two light chains with a stoichiometry of 1.8:1.8 and mol wt of 20,000 and 16,000 daltons. Clearly, the faster migrating light chain, LC3, found in standard cultures is synthesized not by the myotubes but ty the mononucleated cells. In myotubes, both the assembly of the sarcomeres and the interaction between thick and thin filaments required for spontaneous contraction occur in the absence of light chain LC3. One set of structural genes for the myosin light and heavy chains appears to be active in mononucleated cells, whereas another set appears to be active in multinucleated myotubes.

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Selected References

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  1. Abbott J., Schiltz J., Dienstman S., Holtzer H. The phenotypic complexity of myogenic clones. Proc Natl Acad Sci U S A. 1974 Apr;71(4):1506–1510. doi: 10.1073/pnas.71.4.1506. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Adelman M. R., Taylor E. W. Isolation of an actomyosin-like protein complex from slime mold plasmodium and the separation of the complex into actin- and myosin-like fractions. Biochemistry. 1969 Dec;8(12):4964–4975. doi: 10.1021/bi00840a046. [DOI] [PubMed] [Google Scholar]
  3. Adelstein R. S., Conti M. A., Johnson G. S., Pastan I., Pollard T. D. Isolation and characterization of myosin from cloned mouse fibroblasts. Proc Natl Acad Sci U S A. 1972 Dec;69(12):3693–3697. doi: 10.1073/pnas.69.12.3693. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Adelstein R. S., Pollard T. D., Kuehl W. M. Isolation and characterization of myosin and two myosin fragments from human blood platelets. Proc Natl Acad Sci U S A. 1971 Nov;68(11):2703–2707. doi: 10.1073/pnas.68.11.2703. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Holtzer H., Strahs K., Biehl J., Somlyo A. P., Ishikawa H. Thick and thin filaments in postmitotic, mononucleated myoblasts. Science. 1975 May 30;188(4191):943–945. doi: 10.1126/science.1138363. [DOI] [PubMed] [Google Scholar]
  6. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  7. Morimoto K., Harrington W. F. Isolation and composition of thick filaments from rabbit skeletal muscle. J Mol Biol. 1973 Jun 15;77(1):165–175. doi: 10.1016/0022-2836(73)90370-7. [DOI] [PubMed] [Google Scholar]
  8. Okazaki K., Holtzer H. Myogenesis: fusion, myosin synthesis, and the mitotic cycle. Proc Natl Acad Sci U S A. 1966 Nov;56(5):1484–1490. doi: 10.1073/pnas.56.5.1484. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Orkin R. W., Pollard T. D., Hay E. D. SDS gel analysis of muscle proteins in embryonic cells. Dev Biol. 1973 Dec;35(2):388–394. doi: 10.1016/0012-1606(73)90035-3. [DOI] [PubMed] [Google Scholar]
  10. Paterson B., Strohman R. C. Myosin synthesis in cultures of differentiating chicken embryo skeletal muscle. Dev Biol. 1972 Oct;29(2):113–138. doi: 10.1016/0012-1606(72)90050-4. [DOI] [PubMed] [Google Scholar]
  11. Pollard T. D., Thomas S. M., Niederman R. Human platelet myosin. I. Purification by a rapid method applicable to other nonmuscle cells. Anal Biochem. 1974 Jul;60(1):258–266. doi: 10.1016/0003-2697(74)90152-3. [DOI] [PubMed] [Google Scholar]
  12. Prives J. M., Paterson B. M. Differentiation of cell membranes in cultures of embryonic chick breast muscle. Proc Natl Acad Sci U S A. 1974 Aug;71(8):3208–3211. doi: 10.1073/pnas.71.8.3208. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Przybyla A., Strohman R. C. Myosin heavy chain messenger RNA from myogenic cell cultures. Proc Natl Acad Sci U S A. 1974 Mar;71(3):662–666. doi: 10.1073/pnas.71.3.662. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Rubinstein N. A., Chi J. C., Holtzer H. Actin and myosin in a variety of myogenic and non-myogenic cells. Biochem Biophys Res Commun. 1974 Mar 25;57(2):438–446. doi: 10.1016/0006-291x(74)90950-4. [DOI] [PubMed] [Google Scholar]
  15. Sarkar S., Sreter F. A., Gergely J. Light chains of myosins from white, red, and cardiac muscles. Proc Natl Acad Sci U S A. 1971 May;68(5):946–950. doi: 10.1073/pnas.68.5.946. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Schroeder T. E. Actin in dividing cells: contractile ring filaments bind heavy meromyosin. Proc Natl Acad Sci U S A. 1973 Jun;70(6):1688–1692. doi: 10.1073/pnas.70.6.1688. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Sreter F., Holtzer S., Gergely J., Holtzer H. Some properties of embryonic myosin. J Cell Biol. 1972 Dec;55(3):586–594. doi: 10.1083/jcb.55.3.586. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Tarikas H., Schubert D. Regulation of adenylate kinase and creatine kinase activities in myogenic cells. Proc Natl Acad Sci U S A. 1974 Jun;71(6):2377–2381. doi: 10.1073/pnas.71.6.2377. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Tilney L. G., Mooseker M. Actin in the brush-border of epithelial cells of the chicken intestine. Proc Natl Acad Sci U S A. 1971 Oct;68(10):2611–2615. doi: 10.1073/pnas.68.10.2611. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Tregear R. T., Squire J. M. Myosin content and filament structure in smooth and striated muscle. J Mol Biol. 1973 Jun 25;77(2):279–290. doi: 10.1016/0022-2836(73)90336-7. [DOI] [PubMed] [Google Scholar]
  21. Wachsberger P. R., Pepe F. A. Purification of uterine myosin and synthetic filament formation. J Mol Biol. 1974 Sep 15;88(2):385–391. doi: 10.1016/0022-2836(74)90489-6. [DOI] [PubMed] [Google Scholar]
  22. Weber K., Groeschel-Stewart U. Antibody to myosin: the specific visualization of myosin-containing filaments in nonmuscle cells. Proc Natl Acad Sci U S A. 1974 Nov;71(11):4561–4564. doi: 10.1073/pnas.71.11.4561. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Weber K., Osborn M. The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis. J Biol Chem. 1969 Aug 25;244(16):4406–4412. [PubMed] [Google Scholar]
  24. Weeds A. G., Hall R., Spurway N. C. Characterization of myosin light chains from histochemically identified fibres of rabbit psoas muscle. FEBS Lett. 1975 Jan 1;49(3):320–324. doi: 10.1016/0014-5793(75)80776-9. [DOI] [PubMed] [Google Scholar]
  25. Weeds A. G. Light chains of myosin. Nature. 1969 Sep 27;223(5213):1362–1364. doi: 10.1038/2231362a0. [DOI] [PubMed] [Google Scholar]
  26. Weeds A. G., Lowey S. Substructure of the myosin molecule. II. The light chains of myosin. J Mol Biol. 1971 Nov 14;61(3):701–725. doi: 10.1016/0022-2836(71)90074-x. [DOI] [PubMed] [Google Scholar]
  27. Willingham M. C., Ostlund R. E., Pastan I. Myosin is a component of the cell surface of cultured cells. Proc Natl Acad Sci U S A. 1974 Oct;71(10):4144–4148. doi: 10.1073/pnas.71.10.4144. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. YAFFE D., FELDMAN M. THE EFFECT OF ACTINOMYCIN D ON HEART AND THIGH MUSCLE CELLS GROWN IN VITRO. Dev Biol. 1964 Jun;9:347–366. doi: 10.1016/0012-1606(64)90030-2. [DOI] [PubMed] [Google Scholar]
  29. Yang Y. Z., Perdue J. F. Contractile proteins of cultured cells. I. The isolation and characterization of an actin-like protein from cultured chick embryo fibroblasts. J Biol Chem. 1972 Jul 25;247(14):4503–4509. [PubMed] [Google Scholar]

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