Skip to main content
Molecular and Cellular Biology logoLink to Molecular and Cellular Biology
. 1989 Aug;9(8):3203–3211. doi: 10.1128/mcb.9.8.3203

Mitogen stimulation affects contractile protein mRNA abundance and translation in embryonic quail myocytes.

K E Latham 1, I R Konigsberg 1
PMCID: PMC362364  PMID: 2796987

Abstract

In cultures of differentiated, fusion-blocked muscle cells obtained from embryonic Japanese quail (Coturnix coturnix japonica), mitogen stimulation leads to an immediate reduction in the rates of synthesis of skeletal muscle myosin heavy chain (MHC) and alpha-actin. The molecular mechanisms responsible for this downregulation were examined. The cellular abundances of the alpha-actin and MHC mRNAs were affected differently by mitogen stimulation; alpha-actin mRNA abundance declined by an amount which quantitatively accounted for the observed decrease in alpha-actin synthesis, whereas MHC mRNA abundance remained virtually unchanged during the first 6 h following mitogen stimulation, a period during which MHC synthesis declined by more than 70%. MHC mRNA abundance did decline between 6 and 12 h after mitogen stimulation. Downregulation of MHC synthesis therefore involves an initial block in mRNA translation combined with a later loss of MHC mRNA from the cytoplasma, while alpha-actin synthesis is regulated at the level of mRNA abundance. These observations are consistent with the hypothesis that, in addition to transcriptional activation of muscle-specific genes, skeletal muscle differentiation normally involves cell cycle-dependent modulations in cellular factors which control message stability and message translation.

Full text

PDF
3203

Images in this article

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. BREUER C. B., DAVIES M. C., FLORINI J. R. AMINO ACID INCORPORATION INTO PROTEIN BY CELL-FREE PREPARATIONS FROM RAT SKELETAL MUSCLE. II. PREPARATION AND PROPERTIES OF MUSCLE RIBOSOMES AND POLYRIBOSOMES. Biochemistry. 1964 Nov;3:1713–1719. doi: 10.1021/bi00899a020. [DOI] [PubMed] [Google Scholar]
  2. Bains W., Ponte P., Blau H., Kedes L. Cardiac actin is the major actin gene product in skeletal muscle cell differentiation in vitro. Mol Cell Biol. 1984 Aug;4(8):1449–1453. doi: 10.1128/mcb.4.8.1449. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bayne E. K., Simpson S. B., Jr Influence of environmental factors on the accumulation and differentiation of prefusion G1 lizard myoblasts in vitro. Exp Cell Res. 1980 May;127(1):15–30. doi: 10.1016/0014-4827(80)90411-5. [DOI] [PubMed] [Google Scholar]
  4. Berk A. J., Sharp P. A. Sizing and mapping of early adenovirus mRNAs by gel electrophoresis of S1 endonuclease-digested hybrids. Cell. 1977 Nov;12(3):721–732. doi: 10.1016/0092-8674(77)90272-0. [DOI] [PubMed] [Google Scholar]
  5. Bowman L. H., Emerson C. P., Jr Formation and stability of cytoplasmic mRNAs during myoblast differentiation: pulse-chase and density labeling analyses. Dev Biol. 1980 Nov;80(1):146–166. doi: 10.1016/0012-1606(80)90505-9. [DOI] [PubMed] [Google Scholar]
  6. Buckingham M. E., Caput D., Cohen A., Whalen R. G., Gros F. The synthesis and stability of cytoplasmic messenger RNA during myoblast differentiation in culture. Proc Natl Acad Sci U S A. 1974 Apr;71(4):1466–1470. doi: 10.1073/pnas.71.4.1466. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Chen C. Y., Beatty J. T., Cohen S. N., Belasco J. G. An intercistronic stem-loop structure functions as an mRNA decay terminator necessary but insufficient for puf mRNA stability. Cell. 1988 Feb 26;52(4):609–619. doi: 10.1016/0092-8674(88)90473-4. [DOI] [PubMed] [Google Scholar]
  8. Clegg C. H., Linkhart T. A., Olwin B. B., Hauschka S. D. Growth factor control of skeletal muscle differentiation: commitment to terminal differentiation occurs in G1 phase and is repressed by fibroblast growth factor. J Cell Biol. 1987 Aug;105(2):949–956. doi: 10.1083/jcb.105.2.949. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Compton R. S., Konigsberg I. R. Cell cycle withdrawal without concomitant differentiation: analysis of a G1-specific temperature-sensitive murine myoblast cell line. Dev Biol. 1988 Oct;129(2):476–494. doi: 10.1016/0012-1606(88)90394-6. [DOI] [PubMed] [Google Scholar]
  10. Craig N. The effects of inhibitors of RNA and DNA synthesis on protein synthesis and polysome levels in mouse L-cells. J Cell Physiol. 1973 Oct;82(2):133–150. doi: 10.1002/jcp.1040820202. [DOI] [PubMed] [Google Scholar]
  11. Devlin B. H., Konigsberg I. R. Reentry into the cell cycle of differentiated skeletal myocytes. Dev Biol. 1983 Jan;95(1):175–192. doi: 10.1016/0012-1606(83)90016-7. [DOI] [PubMed] [Google Scholar]
  12. Devlin R. B., Emerson C. P., Jr Coordinate accumulation of contractile protein mRNAs during myoblast differentiation. Dev Biol. 1979 Mar;69(1):202–216. doi: 10.1016/0012-1606(79)90286-0. [DOI] [PubMed] [Google Scholar]
  13. Eldridge J., Zehner Z., Paterson B. M. Nucleotide sequence of the chicken cardiac alpha actin gene: absence of strong homologies in the promoter and 3'-untranslated regions with the skeletal alpha actin sequence. Gene. 1985;36(1-2):55–63. doi: 10.1016/0378-1119(85)90069-1. [DOI] [PubMed] [Google Scholar]
  14. Emerson C. P., Jr, Beckner S. K. Activation of myosin synthesis in fusing and mononucleated myoblasts. J Mol Biol. 1975 Apr 25;93(4):431–447. doi: 10.1016/0022-2836(75)90238-7. [DOI] [PubMed] [Google Scholar]
  15. Endo T., Nadal-Ginard B. Three types of muscle-specific gene expression in fusion-blocked rat skeletal muscle cells: translational control in EGTA-treated cells. Cell. 1987 May 22;49(4):515–526. doi: 10.1016/0092-8674(87)90454-5. [DOI] [PubMed] [Google Scholar]
  16. Feinberg A. P., Vogelstein B. "A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity". Addendum. Anal Biochem. 1984 Feb;137(1):266–267. doi: 10.1016/0003-2697(84)90381-6. [DOI] [PubMed] [Google Scholar]
  17. Feinberg A. P., Vogelstein B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem. 1983 Jul 1;132(1):6–13. doi: 10.1016/0003-2697(83)90418-9. [DOI] [PubMed] [Google Scholar]
  18. Florini J. R., Roberts A. B., Ewton D. Z., Falen S. L., Flanders K. C., Sporn M. B. Transforming growth factor-beta. A very potent inhibitor of myoblast differentiation, identical to the differentiation inhibitor secreted by Buffalo rat liver cells. J Biol Chem. 1986 Dec 15;261(35):16509–16513. [PubMed] [Google Scholar]
  19. Gospodarowicz D., Weseman J., Moran J. S., Lindstrom J. Effect of fibroblast growth factor on the division and fusion of bovine myoblasts. J Cell Biol. 1976 Aug;70(2 Pt 1):395–405. doi: 10.1083/jcb.70.2.395. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Hastings K. E., Emerson C. P., Jr cDNA clone analysis of six co-regulated mRNAs encoding skeletal muscle contractile proteins. Proc Natl Acad Sci U S A. 1982 Mar;79(5):1553–1557. doi: 10.1073/pnas.79.5.1553. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Hayward L. J., Schwartz R. J. Sequential expression of chicken actin genes during myogenesis. J Cell Biol. 1986 Apr;102(4):1485–1493. doi: 10.1083/jcb.102.4.1485. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Heywood S. M., Dowben R. M., Rich A. A study of muscle polyribosomes and the coprecipitation of polyribosomes with myosin. Biochemistry. 1968 Sep;7(9):3289–3296. doi: 10.1021/bi00849a036. [DOI] [PubMed] [Google Scholar]
  23. Heywood S. M., Nwagwu M. Partial characterization of presumptive myosin messenger ribonucleic acid. Biochemistry. 1969 Sep;8(9):3839–3845. doi: 10.1021/bi00837a050. [DOI] [PubMed] [Google Scholar]
  24. Konieczny S. F., Emerson C. P., Jr Differentiation, not determination, regulates muscle gene activation: transfection of troponin I genes into multipotential and muscle lineages of 10T1/2 cells. Mol Cell Biol. 1985 Sep;5(9):2423–2432. doi: 10.1128/mcb.5.9.2423. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Konigsberg I. R. Diffusion-mediated control of myoblast fusion. Dev Biol. 1971 Sep;26(1):133–152. doi: 10.1016/0012-1606(71)90113-8. [DOI] [PubMed] [Google Scholar]
  26. Lathrop B., Olson E., Glaser L. Control by fibroblast growth factor of differentiation in the BC3H1 muscle cell line. J Cell Biol. 1985 May;100(5):1540–1547. doi: 10.1083/jcb.100.5.1540. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Lathrop B., Thomas K., Glaser L. Control of myogenic differentiation by fibroblast growth factor is mediated by position in the G1 phase of the cell cycle. J Cell Biol. 1985 Dec;101(6):2194–2198. doi: 10.1083/jcb.101.6.2194. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Linkhart T. A., Clegg C. H., Hauschika S. D. Myogenic differentiation in permanent clonal mouse myoblast cell lines: regulation by macromolecular growth factors in the culture medium. Dev Biol. 1981 Aug;86(1):19–30. doi: 10.1016/0012-1606(81)90311-0. [DOI] [PubMed] [Google Scholar]
  29. Linkhart T. A., Clegg C. H., Hauschka S. D. Control of mouse myoblast commitment to terminal differentiation by mitogens. J Supramol Struct. 1980;14(4):483–498. doi: 10.1002/jss.400140407. [DOI] [PubMed] [Google Scholar]
  30. Magnaldo I., L'Allemain G., Chambard J. C., Moenner M., Barritault D., Pouysségur J. The mitogenic signaling pathway of fibroblast growth factor is not mediated through polyphosphoinositide hydrolysis and protein kinase C activation in hamster fibroblasts. J Biol Chem. 1986 Dec 25;261(36):16916–16922. [PubMed] [Google Scholar]
  31. Massagué J., Cheifetz S., Endo T., Nadal-Ginard B. Type beta transforming growth factor is an inhibitor of myogenic differentiation. Proc Natl Acad Sci U S A. 1986 Nov;83(21):8206–8210. doi: 10.1073/pnas.83.21.8206. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Müllner E. W., Kühn L. C. A stem-loop in the 3' untranslated region mediates iron-dependent regulation of transferrin receptor mRNA stability in the cytoplasm. Cell. 1988 Jun 3;53(5):815–825. doi: 10.1016/0092-8674(88)90098-0. [DOI] [PubMed] [Google Scholar]
  33. Nadal-Ginard B. Commitment, fusion and biochemical differentiation of a myogenic cell line in the absence of DNA synthesis. Cell. 1978 Nov;15(3):855–864. doi: 10.1016/0092-8674(78)90270-2. [DOI] [PubMed] [Google Scholar]
  34. Nguyen H. T., Medford R. M., Nadal-Ginard B. Reversibility of muscle differentiation in the absence of commitment: analysis of a myogenic cell line temperature-sensitive for commitment. Cell. 1983 Aug;34(1):281–293. doi: 10.1016/0092-8674(83)90159-9. [DOI] [PubMed] [Google Scholar]
  35. O'Farrell P. H. High resolution two-dimensional electrophoresis of proteins. J Biol Chem. 1975 May 25;250(10):4007–4021. [PMC free article] [PubMed] [Google Scholar]
  36. Olson E. N., Caldwell K. L., Gordon J. I., Glaser L. Regulation of creatine phosphokinase expression during differentiation of BC3H1 cells. J Biol Chem. 1983 Feb 25;258(4):2644–2652. [PubMed] [Google Scholar]
  37. Olson E. N., Sternberg E., Hu J. S., Spizz G., Wilcox C. Regulation of myogenic differentiation by type beta transforming growth factor. J Cell Biol. 1986 Nov;103(5):1799–1805. doi: 10.1083/jcb.103.5.1799. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Paterson B. M., Eldridge J. D. alpha-Cardiac actin is the major sarcomeric isoform expressed in embryonic avian skeletal muscle. Science. 1984 Jun 29;224(4656):1436–1438. doi: 10.1126/science.6729461. [DOI] [PubMed] [Google Scholar]
  39. Pontecorvi A., Tata J. R., Phyillaier M., Robbins J. Selective degradation of mRNA: the role of short-lived proteins in differential destabilization of insulin-induced creatine phosphokinase and myosin heavy chain mRNAs during rat skeletal muscle L6 cell differentiation. EMBO J. 1988 May;7(5):1489–1495. doi: 10.1002/j.1460-2075.1988.tb02967.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Shaw G., Kamen R. A conserved AU sequence from the 3' untranslated region of GM-CSF mRNA mediates selective mRNA degradation. Cell. 1986 Aug 29;46(5):659–667. doi: 10.1016/0092-8674(86)90341-7. [DOI] [PubMed] [Google Scholar]
  41. Slater C. R. Control of myogenesis in vitro by chick embryo extract. Dev Biol. 1976 Jun;50(2):264–284. doi: 10.1016/0012-1606(76)90151-2. [DOI] [PubMed] [Google Scholar]
  42. Spizz G., Roman D., Strauss A., Olson E. N. Serum and fibroblast growth factor inhibit myogenic differentiation through a mechanism dependent on protein synthesis and independent of cell proliferation. J Biol Chem. 1986 Jul 15;261(20):9483–9488. [PubMed] [Google Scholar]
  43. Umeda P. K., Sinha A. M., Jakovcic S., Merten S., Hsu H. J., Subramanian K. N., Zak R., Rabinowitz M. Molecular cloning of two fast myosin heavy chain cDNAs from chicken embryo skeletal muscle. Proc Natl Acad Sci U S A. 1981 May;78(5):2843–2847. doi: 10.1073/pnas.78.5.2843. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Vieira J., Messing J. The pUC plasmids, an M13mp7-derived system for insertion mutagenesis and sequencing with synthetic universal primers. Gene. 1982 Oct;19(3):259–268. doi: 10.1016/0378-1119(82)90015-4. [DOI] [PubMed] [Google Scholar]

Articles from Molecular and Cellular Biology are provided here courtesy of Taylor & Francis

RESOURCES