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. 1986 Jun;6(6):2137–2148. doi: 10.1128/mcb.6.6.2137

Two-level regulation of cardiac actin gene transcription: muscle-specific modulating factors can accumulate before gene activation.

A Minty, H Blau, L Kedes
PMCID: PMC367754  PMID: 3785190

Abstract

We have previously proposed that the upstream regions of the human cardiac actin gene contain sequences that interact with muscle-specific factors with direct high-level transcription of this gene in differentiated muscle cells. In this study we showed that these factors already accumulate in the dividing myoblasts of the mouse C2C12 cell line before differentiation of the cells. The endogenous cardiac actin gene in the C2C12 line is expressed only at a low level in myoblasts but at a high level when these cells differentiate into multinucleate myotubes. In contrast, human cardiac actin genes stably introduced into C2C12 cells show high-level expression in both myoblasts and myotubes, indicating that the endogenous cardiac actin gene is repressed in myoblasts by a mechanism which does not affect transfected genes. In a second muscle cell line (the rat L8 cell line), the level of expression of transfected cardiac actin genes increases when these cells differentiate into myotubes, paralleling the expression of the endogenous sarcomeric actin genes. We suggest that the level of transcriptional modulating factors is low in L8 myoblasts and increases when these cells differentiate into myotubes. Our results demonstrate that at least two steps are necessary for high-level cardiac actin gene expression: activation of the gene and subsequent modulation of its transcriptional activity. Furthermore, the results indicate that the two regulatory steps can be dissociated and that the factors involved in modulation are distinct from those involved in gene activation.

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Selected References

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  1. Alonso S., Minty A., Bourlet Y., Buckingham M. Comparison of three actin-coding sequences in the mouse; evolutionary relationships between the actin genes of warm-blooded vertebrates. J Mol Evol. 1986;23(1):11–22. doi: 10.1007/BF02100994. [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. Blau H. M., Chiu C. P., Webster C. Cytoplasmic activation of human nuclear genes in stable heterocaryons. Cell. 1983 Apr;32(4):1171–1180. doi: 10.1016/0092-8674(83)90300-8. [DOI] [PubMed] [Google Scholar]
  4. Blau H. M., Epstein C. J. Manipulation of myogenesis in vitro: reversible inhibition by DMSO. Cell. 1979 May;17(1):95–108. doi: 10.1016/0092-8674(79)90298-8. [DOI] [PubMed] [Google Scholar]
  5. Blau H. M., Pavlath G. K., Hardeman E. C., Chiu C. P., Silberstein L., Webster S. G., Miller S. C., Webster C. Plasticity of the differentiated state. Science. 1985 Nov 15;230(4727):758–766. doi: 10.1126/science.2414846. [DOI] [PubMed] [Google Scholar]
  6. Borrelli E., Hen R., Chambon P. Adenovirus-2 E1A products repress enhancer-induced stimulation of transcription. Nature. 1984 Dec 13;312(5995):608–612. doi: 10.1038/312608a0. [DOI] [PubMed] [Google Scholar]
  7. Brown D. D. The role of stable complexes that repress and activate eucaryotic genes. Cell. 1984 Jun;37(2):359–365. doi: 10.1016/0092-8674(84)90366-0. [DOI] [PubMed] [Google Scholar]
  8. Caravatti M., Minty A., Robert B., Montarras D., Weydert A., Cohen A., Daubas P., Buckingham M. Regulation of muscle gene expression. The accumulation of messenger RNAs coding for muscle-specific proteins during myogenesis in a mouse cell line. J Mol Biol. 1982 Sep;160(1):59–76. doi: 10.1016/0022-2836(82)90131-0. [DOI] [PubMed] [Google Scholar]
  9. Carmon Y., Czosnek H., Nudel U., Shani M., Yaffe D. DNAase I sensitivity of genes expressed during myogenesis. Nucleic Acids Res. 1982 May 25;10(10):3085–3098. doi: 10.1093/nar/10.10.3085. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Charnay P., Treisman R., Mellon P., Chao M., Axel R., Maniatis T. Differences in human alpha- and beta-globin gene expression in mouse erythroleukemia cells: the role of intragenic sequences. Cell. 1984 Aug;38(1):251–263. doi: 10.1016/0092-8674(84)90547-6. [DOI] [PubMed] [Google Scholar]
  11. Chiu C. P., Blau H. M. 5-Azacytidine permits gene activation in a previously noninducible cell type. Cell. 1985 Feb;40(2):417–424. doi: 10.1016/0092-8674(85)90155-2. [DOI] [PubMed] [Google Scholar]
  12. Davies R. L., Fuhrer-Krusi S., Kucherlapati R. S. Modulation of transfected gene expression mediated by changes in chromatin structure. Cell. 1982 Dec;31(3 Pt 2):521–529. doi: 10.1016/0092-8674(82)90308-7. [DOI] [PubMed] [Google Scholar]
  13. Engel J., Gunning P., Kedes L. Human cytoplasmic actin proteins are encoded by a multigene family. Mol Cell Biol. 1982 Jun;2(6):674–684. doi: 10.1128/mcb.2.6.674. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Gilmour R. S., Spandidos D. A., Vass J. K., Gow J. W., Paul J. A negative regulatory sequence near the mouse beta-maj globin gene associated with a region of potential Z-DNA. EMBO J. 1984 Jun;3(6):1263–1272. doi: 10.1002/j.1460-2075.1984.tb01961.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Gorman C. M., Rigby P. W., Lane D. P. Negative regulation of viral enhancers in undifferentiated embryonic stem cells. Cell. 1985 Sep;42(2):519–526. doi: 10.1016/0092-8674(85)90109-6. [DOI] [PubMed] [Google Scholar]
  16. Groudine M., Casimir C. Post-transcriptional regulation of the chicken thymidine kinase gene. Nucleic Acids Res. 1984 Feb 10;12(3):1427–1446. doi: 10.1093/nar/12.3.1427. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Gunning P., Mohun T., Ng S. Y., Ponte P., Kedes L. Evolution of the human sarcomeric-actin genes: evidence for units of selection within the 3' untranslated regions of the mRNAs. J Mol Evol. 1984;20(3-4):202–214. doi: 10.1007/BF02104727. [DOI] [PubMed] [Google Scholar]
  18. Gunning P., Ponte P., Blau H., Kedes L. alpha-skeletal and alpha-cardiac actin genes are coexpressed in adult human skeletal muscle and heart. Mol Cell Biol. 1983 Nov;3(11):1985–1995. doi: 10.1128/mcb.3.11.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Gunning P., Ponte P., Kedes L., Hickey R. J., Skoultchi A. I. Expression of human cardiac actin in mouse L cells: a sarcomeric actin associates with a nonmuscle cytoskeleton. Cell. 1984 Mar;36(3):709–715. doi: 10.1016/0092-8674(84)90351-9. [DOI] [PubMed] [Google Scholar]
  20. Gurdon J. B., Brennan S., Fairman S., Mohun T. J. Transcription of muscle-specific actin genes in early Xenopus development: nuclear transplantation and cell dissociation. Cell. 1984 Oct;38(3):691–700. doi: 10.1016/0092-8674(84)90264-2. [DOI] [PubMed] [Google Scholar]
  21. 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]
  22. Inestrosa N. C., Miller J. B., Silberstein L., Ziskind-Conhaim L., Hall Z. W. Developmental regulation of 16S acetylcholinesterase and acetylcholine receptors in a mouse muscle cell line. Exp Cell Res. 1983 Sep;147(2):393–405. doi: 10.1016/0014-4827(83)90221-5. [DOI] [PubMed] [Google Scholar]
  23. Jones P. B., Galeazzi D. R., Fisher J. M., Whitlock J. P., Jr Control of cytochrome P1-450 gene expression by dioxin. Science. 1985 Mar 22;227(4693):1499–1502. doi: 10.1126/science.3856321. [DOI] [PubMed] [Google Scholar]
  24. Keshet I., Yisraeli J., Cedar H. Effect of regional DNA methylation on gene expression. Proc Natl Acad Sci U S A. 1985 May;82(9):2560–2564. doi: 10.1073/pnas.82.9.2560. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Killary A. M., Fournier R. E. A genetic analysis of extinction: trans-dominant loci regulate expression of liver-specific traits in hepatoma hybrid cells. Cell. 1984 Sep;38(2):523–534. doi: 10.1016/0092-8674(84)90507-5. [DOI] [PubMed] [Google Scholar]
  26. 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]
  27. Mayer Y., Czosnek H., Zeelon P. E., Yaffe D., Nudel U. Expression of the genes coding for the skeletal muscle and cardiac actions in the heart. Nucleic Acids Res. 1984 Jan 25;12(2):1087–1100. doi: 10.1093/nar/12.2.1087. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. McKenna N., Meigs J. B., Wang Y. L. Identical distribution of fluorescently labeled brain and muscle actins in living cardiac fibroblasts and myocytes. J Cell Biol. 1985 Jan;100(1):292–296. doi: 10.1083/jcb.100.1.292. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Medford R. M., Nguyen H. T., Nadal-Ginard B. Transcriptional and cell cycle-mediated regulation of myosin heavy chain gene expression during muscle cell differentiation. J Biol Chem. 1983 Sep 25;258(18):11063–11073. [PubMed] [Google Scholar]
  30. Melloul D., Aloni B., Calvo J., Yaffe D., Nudel U. Developmentally regulated expression of chimeric genes containing muscle actin DNA sequences in transfected myogenic cells. EMBO J. 1984 May;3(5):983–990. doi: 10.1002/j.1460-2075.1984.tb01917.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Merrill G. F., Hauschka S. D., McKnight S. L. tk Enzyme expression in differentiating muscle cells is regulated through an internal segment of the cellular tk gene. Mol Cell Biol. 1984 Sep;4(9):1777–1784. doi: 10.1128/mcb.4.9.1777. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Minty A. J., Alonso S., Caravatti M., Buckingham M. E. A fetal skeletal muscle actin mRNA in the mouse and its identity with cardiac actin mRNA. Cell. 1982 Aug;30(1):185–192. doi: 10.1016/0092-8674(82)90024-1. [DOI] [PubMed] [Google Scholar]
  33. Minty A. J., Gros F. Coding potential of non-polyadenylated messenger RNA in mouse Friend cells. J Mol Biol. 1980 May 5;139(1):61–83. doi: 10.1016/0022-2836(80)90116-3. [DOI] [PubMed] [Google Scholar]
  34. Minty A., Blau H., Kedes L. Two-level regulation of cardiac actin gene transcription: muscle-specific modulating factors can accumulate before gene activation. Mol Cell Biol. 1986 Jun;6(6):2137–2148. doi: 10.1128/mcb.6.6.2137. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Mohun T. J., Brennan S., Dathan N., Fairman S., Gurdon J. B. Cell type-specific activation of actin genes in the early amphibian embryo. Nature. 1984 Oct 25;311(5988):716–721. doi: 10.1038/311716a0. [DOI] [PubMed] [Google Scholar]
  36. 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]
  37. Nudel U., Greenberg D., Ordahl C. P., Saxel O., Neuman S., Yaffe D. Developmentally regulated expression of a chicken muscle-specific gene in stably transfected rat myogenic cells. Proc Natl Acad Sci U S A. 1985 May;82(10):3106–3109. doi: 10.1073/pnas.82.10.3106. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Ornitz D. M., Palmiter R. D., Hammer R. E., Brinster R. L., Swift G. H., MacDonald R. J. Specific expression of an elastase-human growth hormone fusion gene in pancreatic acinar cells of transgenic mice. Nature. 1985 Feb 14;313(6003):600–602. doi: 10.1038/313600a0. [DOI] [PubMed] [Google Scholar]
  39. 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]
  40. Robins D. M., Paek I., Seeburg P. H., Axel R. Regulated expression of human growth hormone genes in mouse cells. Cell. 1982 Jun;29(2):623–631. doi: 10.1016/0092-8674(82)90178-7. [DOI] [PubMed] [Google Scholar]
  41. Scott R. W., Vogt T. F., Croke M. E., Tilghman S. M. Tissue-specific activation of a cloned alpha-fetoprotein gene during differentiation of a transfected embryonal carcinoma cell line. Nature. 1984 Aug 16;310(5978):562–567. doi: 10.1038/310562a0. [DOI] [PubMed] [Google Scholar]
  42. Seiler-Tuyns A., Eldridge J. D., Paterson B. M. Expression and regulation of chicken actin genes introduced into mouse myogenic and nonmyogenic cells. Proc Natl Acad Sci U S A. 1984 May;81(10):2980–2984. doi: 10.1073/pnas.81.10.2980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Shani M., Admon S., Yaffe D. The methylation state of 2 muscle-specific genes: restriction enzyme analysis did not detect a correlation with expression. Nucleic Acids Res. 1984 Sep 25;12(18):7225–7234. doi: 10.1093/nar/12.18.7225. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Shani M. Tissue-specific expression of rat myosin light-chain 2 gene in transgenic mice. Nature. 1985 Mar 21;314(6008):283–286. doi: 10.1038/314283a0. [DOI] [PubMed] [Google Scholar]
  45. Shani M., Zevin-Sonkin D., Saxel O., Carmon Y., Katcoff D., Nudel U., Yaffe D. The correlation between the synthesis of skeletal muscle actin, myosin heavy chain, and myosin light chain and the accumulation of corresponding mRNA sequences during myogenesis. Dev Biol. 1981 Sep;86(2):483–492. doi: 10.1016/0012-1606(81)90206-2. [DOI] [PubMed] [Google Scholar]
  46. Shaw P., Sordat B., Schibler U. The two promoters of the mouse alpha-amylase gene Amy-1a are differentially activated during parotid gland differentiation. Cell. 1985 Apr;40(4):907–912. doi: 10.1016/0092-8674(85)90350-2. [DOI] [PubMed] [Google Scholar]
  47. Southern P. J., Berg P. Transformation of mammalian cells to antibiotic resistance with a bacterial gene under control of the SV40 early region promoter. J Mol Appl Genet. 1982;1(4):327–341. [PubMed] [Google Scholar]
  48. Vandekerckhove J., Weber K. Chordate muscle actins differ distinctly from invertebrate muscle actins. The evolution of the different vertebrate muscle actins. J Mol Biol. 1984 Nov 5;179(3):391–413. doi: 10.1016/0022-2836(84)90072-x. [DOI] [PubMed] [Google Scholar]
  49. Wasylyk B., Wasylyk C., Chambon P. Short and long range activation by the SV40 enhancer. Nucleic Acids Res. 1984 Jul 25;12(14):5589–5608. doi: 10.1093/nar/12.14.5589. [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. Weintraub H. A dominant role for DNA secondary structure in forming hypersensitive structures in chromatin. Cell. 1983 Apr;32(4):1191–1203. doi: 10.1016/0092-8674(83)90302-1. [DOI] [PubMed] [Google Scholar]
  51. Weintraub H. Assembly and propagation of repressed and depressed chromosomal states. Cell. 1985 Oct;42(3):705–711. doi: 10.1016/0092-8674(85)90267-3. [DOI] [PubMed] [Google Scholar]
  52. Yaffe D., Saxel O. A myogenic cell line with altered serum requirements for differentiation. Differentiation. 1977;7(3):159–166. doi: 10.1111/j.1432-0436.1977.tb01507.x. [DOI] [PubMed] [Google Scholar]
  53. Yaffe D., Saxel O. Serial passaging and differentiation of myogenic cells isolated from dystrophic mouse muscle. Nature. 1977 Dec 22;270(5639):725–727. doi: 10.1038/270725a0. [DOI] [PubMed] [Google Scholar]

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