Skip to main content
PMC home page
The Journal of Clinical Investigation logoLink to The Journal of Clinical Investigation
. 1988 Aug;82(2):524–531. doi: 10.1172/JCI113627

Molecular cloning and characterization of human cardiac alpha- and beta-form myosin heavy chain complementary DNA clones. Regulation of expression during development and pressure overload in human atrium.

M Kurabayashi 1, H Tsuchimochi 1, I Komuro 1, F Takaku 1, Y Yazaki 1
PMCID: PMC303543  PMID: 2969919

Abstract

We have constructed and characterized two types of myosin heavy chain (MHC) cDNA clones (pHMHC2, pHMHC5) from a fetal human heart cDNA library. Comparison of the nucleotide and deduced amino acid sequences between pHMHC2 and pHMHC5 shows 95.1 and 96.2% homology, respectively. The carboxyl-terminal peptide and 3'-untranslated (3'-UT) regions are highly divergent and specific for these cDNA clones. By using the synthetic oligonucleotide probes that are complementary to the unique 3'-UT regions of these cDNA clones, we demonstrate that pHMHC2 is exclusively transcribed in the atrium, whereas the mRNA for pHMHC5 is predominantly expressed in the ventricle. This result indicates that pHMHC2 and pHMHC5 code for alpha- and beta-form MHCs, respectively. Furthermore, we show that beta-form MHC mRNA is expressed in adult atrium at a low level but scarcely expressed in fetal atrium. Finally, we demonstrate that MHC isozymic transition in pressure-overloaded atrium is, at least in part, regulated at a pretranslational level.

Full text

PDF
527

Images in this article

527Image
on p.527
527Image
on p.527
528Image
on p.528
528Image
on p.528
528Image
on p.528
528Image
on p.528
529Image
on p.529

Selected References

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

  1. Chizzonite R. A., Everett A. W., Clark W. A., Jakovcic S., Rabinowitz M., Zak R. Isolation and characterization of two molecular variants of myosin heavy chain from rabbit ventricle. Change in their content during normal growth and after treatment with thyroid hormone. J Biol Chem. 1982 Feb 25;257(4):2056–2065. [PubMed] [Google Scholar]
  2. Clark W. A., Jr, Chizzonite R. A., Everett A. W., Rabinowitz M., Zak R. Species correlations between cardiac isomyosins. A comparison of electrophoretic and immunological properties. J Biol Chem. 1982 May 25;257(10):5449–5454. [PubMed] [Google Scholar]
  3. Cummins P. Transitions in human atrial and ventricular myosin light-chain isoenzymes in response to cardiac-pressure-overload-induced hypertrophy. Biochem J. 1982 Jul 1;205(1):195–204. doi: 10.1042/bj2050195. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Dillmann W. H. Influence of thyroid hormone administration on myosin ATPase activity and myosin isoenzyme distribution in the heart of diabetic rats. Metabolism. 1982 Mar;31(3):199–204. doi: 10.1016/0026-0495(82)90052-x. [DOI] [PubMed] [Google Scholar]
  5. Everett A. W., Clark W. A., Chizzonite R. A., Zak R. Change in synthesis rates of alpha- and beta-myosin heavy chains in rabbit heart after treatment with thyroid hormone. J Biol Chem. 1983 Feb 25;258(4):2421–2425. [PubMed] [Google Scholar]
  6. Flink I. L., Rader J. H., Morkin E. Thyroid hormone stimulates synthesis of a cardiac myosin isozyme. Comparison of the two-two-dimensional electrophoretic patterns of the cyanogen bromide peptides of cardiac myosin heavy chains from euthyroid and thyrotoxic rabbits. J Biol Chem. 1979 Apr 25;254(8):3105–3110. [PubMed] [Google Scholar]
  7. Friedman D. J., Umeda P. K., Sinha A. M., Hsu H. J., Jakovcic S., Rabinowitz M. Characterization of genomic clones specifying rabbit alpha- and beta-ventricular myosin heavy chains. Proc Natl Acad Sci U S A. 1984 May;81(10):3044–3048. doi: 10.1073/pnas.81.10.3044. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Gorza L., Mercadier J. J., Schwartz K., Thornell L. E., Sartore S., Schiaffino S. Myosin types in the human heart. An immunofluorescence study of normal and hypertrophied atrial and ventricular myocardium. Circ Res. 1984 Jun;54(6):694–702. doi: 10.1161/01.res.54.6.694. [DOI] [PubMed] [Google Scholar]
  9. Gubler U., Hoffman B. J. A simple and very efficient method for generating cDNA libraries. Gene. 1983 Nov;25(2-3):263–269. doi: 10.1016/0378-1119(83)90230-5. [DOI] [PubMed] [Google Scholar]
  10. Gupta K. C., Kingsbury D. W. Complete sequences of the intergenic and mRNA start signals in the Sendai virus genome: homologies with the genome of vesicular stomatitis virus. Nucleic Acids Res. 1984 May 11;12(9):3829–3841. doi: 10.1093/nar/12.9.3829. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Gustafson T. A., Markham B. E., Bahl J. J., Morkin E. Thyroid hormone regulates expression of a transfected alpha-myosin heavy-chain fusion gene in fetal heart cells. Proc Natl Acad Sci U S A. 1987 May;84(10):3122–3126. doi: 10.1073/pnas.84.10.3122. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Hoh J. F., Egerton L. J. Action of triiodothyronine on the synthesis of rat ventricular myosin isoenzymes. FEBS Lett. 1979 May 1;101(1):143–148. doi: 10.1016/0014-5793(79)81313-7. [DOI] [PubMed] [Google Scholar]
  13. Hoh J. F., McGrath P. A., Hale P. T. Electrophoretic analysis of multiple forms of rat cardiac myosin: effects of hypophysectomy and thyroxine replacement. J Mol Cell Cardiol. 1978 Nov;10(11):1053–1076. doi: 10.1016/0022-2828(78)90401-7. [DOI] [PubMed] [Google Scholar]
  14. Izumo S., Lompré A. M., Matsuoka R., Koren G., Schwartz K., Nadal-Ginard B., Mahdavi V. Myosin heavy chain messenger RNA and protein isoform transitions during cardiac hypertrophy. Interaction between hemodynamic and thyroid hormone-induced signals. J Clin Invest. 1987 Mar;79(3):970–977. doi: 10.1172/JCI112908. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Izumo S., Nadal-Ginard B., Mahdavi V. All members of the MHC multigene family respond to thyroid hormone in a highly tissue-specific manner. Science. 1986 Feb 7;231(4738):597–600. doi: 10.1126/science.3945800. [DOI] [PubMed] [Google Scholar]
  16. Jandreski M. A., Liew C. C. Construction of a human ventricular cDNA library and characterization of a beta myosin heavy chain cDNA clone. Hum Genet. 1987 May;76(1):47–53. doi: 10.1007/BF00283049. [DOI] [PubMed] [Google Scholar]
  17. Komuro I., Tsuchimochi H., Ueda S., Kurabayashi M., Seko Y., Takaku F., Yazaki Y. Isolation and characterization of two isozymes of myosin heavy chain from canine atrium. J Biol Chem. 1986 Apr 5;261(10):4504–4509. [PubMed] [Google Scholar]
  18. Lehrach H., Diamond D., Wozney J. M., Boedtker H. RNA molecular weight determinations by gel electrophoresis under denaturing conditions, a critical reexamination. Biochemistry. 1977 Oct 18;16(21):4743–4751. doi: 10.1021/bi00640a033. [DOI] [PubMed] [Google Scholar]
  19. Litten R. Z., 3rd, Martin B. J., Low R. B., Alpert N. R. Altered myosin isozyme patterns from pressure-overloaded and thyrotoxic hypertrophied rabbit hearts. Circ Res. 1982 Jun;50(6):856–864. doi: 10.1161/01.res.50.6.856. [DOI] [PubMed] [Google Scholar]
  20. Lompre A. M., Schwartz K., d'Albis A., Lacombe G., Van Thiem N., Swynghedauw B. Myosin isoenzyme redistribution in chronic heart overload. Nature. 1979 Nov 1;282(5734):105–107. doi: 10.1038/282105a0. [DOI] [PubMed] [Google Scholar]
  21. Lompré A. M., Nadal-Ginard B., Mahdavi V. Expression of the cardiac ventricular alpha- and beta-myosin heavy chain genes is developmentally and hormonally regulated. J Biol Chem. 1984 May 25;259(10):6437–6446. [PubMed] [Google Scholar]
  22. Mahdavi V., Chambers A. P., Nadal-Ginard B. Cardiac alpha- and beta-myosin heavy chain genes are organized in tandem. Proc Natl Acad Sci U S A. 1984 May;81(9):2626–2630. doi: 10.1073/pnas.81.9.2626. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Mahdavi V., Periasamy M., Nadal-Ginard B. Molecular characterization of two myosin heavy chain genes expressed in the adult heart. Nature. 1982 Jun 24;297(5868):659–664. doi: 10.1038/297659a0. [DOI] [PubMed] [Google Scholar]
  24. Mercadier J. J., Bouveret P., Gorza L., Schiaffino S., Clark W. A., Zak R., Swynghedauw B., Schwartz K. Myosin isoenzymes in normal and hypertrophied human ventricular myocardium. Circ Res. 1983 Jul;53(1):52–62. doi: 10.1161/01.res.53.1.52. [DOI] [PubMed] [Google Scholar]
  25. Mercadier J. J., Lompré A. M., Wisnewsky C., Samuel J. L., Bercovici J., Swynghedauw B., Schwartz K. Myosin isoenzyme changes in several models of rat cardiac hypertrophy. Circ Res. 1981 Aug;49(2):525–532. doi: 10.1161/01.res.49.2.525. [DOI] [PubMed] [Google Scholar]
  26. Saez L. J., Gianola K. M., McNally E. M., Feghali R., Eddy R., Shows T. B., Leinwand L. A. Human cardiac myosin heavy chain genes and their linkage in the genome. Nucleic Acids Res. 1987 Jul 10;15(13):5443–5459. doi: 10.1093/nar/15.13.5443. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Saez L., Leinwand L. A. Characterization of diverse forms of myosin heavy chain expressed in adult human skeletal muscle. Nucleic Acids Res. 1986 Apr 11;14(7):2951–2969. doi: 10.1093/nar/14.7.2951. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Schier J. J., Adelstein R. S. Structural and enzymatic comparison of human cardiac muscle myosins isolated from infants, adults, and patients with hypertrophic cardiomyopathy. J Clin Invest. 1982 Apr;69(4):816–825. doi: 10.1172/JCI110521. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Sinha A. M., Friedman D. J., Nigro J. M., Jakovcic S., Rabinowitz M., Umeda P. K. Expression of rabbit ventricular alpha-myosin heavy chain messenger RNA sequences in atrial muscle. J Biol Chem. 1984 May 25;259(10):6674–6680. [PubMed] [Google Scholar]
  31. Sinha A. M., Umeda P. K., Kavinsky C. J., Rajamanickam C., Hsu H. J., Jakovcic S., Rabinowitz M. Molecular cloning of mRNA sequences for cardiac alpha- and beta-form myosin heavy chains: expression in ventricles of normal, hypothyroid, and thyrotoxic rabbits. Proc Natl Acad Sci U S A. 1982 Oct;79(19):5847–5851. doi: 10.1073/pnas.79.19.5847. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Tobacman L. S., Adelstein R. S. Enzymatic comparisons between light chain isozymes of human cardiac myosin subfragment-1. J Biol Chem. 1984 Sep 25;259(18):11226–11230. [PubMed] [Google Scholar]
  33. Tsuchimochi H., Sugi M., Kuro-o M., Ueda S., Takaku F., Furuta S., Shirai T., Yazaki Y. Isozymic changes in myosin of human atrial myocardium induced by overload. Immunohistochemical study using monoclonal antibodies. J Clin Invest. 1984 Aug;74(2):662–665. doi: 10.1172/JCI111466. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Weydert A., Daubas P., Lazaridis I., Barton P., Garner I., Leader D. P., Bonhomme F., Catalan J., Simon D., Guénet J. L. Genes for skeletal muscle myosin heavy chains are clustered and are not located on the same mouse chromosome as a cardiac myosin heavy chain gene. Proc Natl Acad Sci U S A. 1985 Nov;82(21):7183–7187. doi: 10.1073/pnas.82.21.7183. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Yazaki Y., Raben M. S. Cardiac myosin adenosinetriphosphatase of rat and mouse. Distinctive enzymatic properties compared with rabbit and dog cardiac myosin. Circ Res. 1974 Jul;35(1):15–23. doi: 10.1161/01.res.35.1.15. [DOI] [PubMed] [Google Scholar]
  36. Yazaki Y., Raben M. S. Effect of the thyroid state on the enzymatic characteristics of cardiac myosin. A difference in behavior of rat and rabbit cardiac myosin. Circ Res. 1975 Jan;36(1):208–215. doi: 10.1161/01.res.36.1.208. [DOI] [PubMed] [Google Scholar]
  37. Yazaki Y., Ueda S., Nagai R., Shimada K. Cardiac atrial myosin adenosine triphosphatase of animals and humans: distinctive enzymatic properties compared with cardiac ventricular myosin. Circ Res. 1979 Oct;45(4):522–527. doi: 10.1161/01.res.45.4.522. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Clinical Investigation are provided here courtesy of American Society for Clinical Investigation

RESOURCES