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. 2000 May 15;348(Pt 1):29–35.

Expression of human hyaluronan synthases in response to external stimuli.

A Jacobson 1, J Brinck 1, M J Briskin 1, A P Spicer 1, P Heldin 1
PMCID: PMC1221032  PMID: 10794710

Abstract

In the present study we have investigated the expression of mRNAs for hyaluronan synthase isoforms (HAS1, HAS2 and HAS3) in different cells in response to various stimuli. Human mesothelial cells, which synthesize large amounts of hyaluronan, express mRNAs encoding all three HAS isoforms, whereas their transformed counterparts, mesothelioma cells, which produce only minute amounts of hyaluronan, express only HAS3 mRNA. Human lung fibroblasts and the glioma cell line U-118 MG express only the HAS2 and HAS3 genes. The expression of the transcripts was higher in subconfluent than in confluent cultures and was well correlated with the production of hyaluronan by the cells. Stimulation of mesothelial cells with platelet-derived growth factor-BB induced an up-regulation of mRNA for HAS2 to a maximum after 6 h of stimulation; HAS1 and HAS3 genes were only induced slightly. Transforming growth factor-beta1 reduced HAS2 mRNA slightly, and hydrocortisone reduced it strongly, within 6 h of stimulation in mesothelial cell cultures but did not significantly affect the expression of mRNAs for HAS1 and HAS3. Induction of HAS1 and HAS2 protein levels in response to the stimuli above correlated with HAS transcript levels. Thus the expression of the three HAS isoforms is more prominent in growing cells than in resting cells and is differentially regulated by various stimuli suggesting distinct functional roles of the three proteins.

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Selected References

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  1. Asplund T., Brinck J., Suzuki M., Briskin M. J., Heldin P. Characterization of hyaluronan synthase from a human glioma cell line. Biochim Biophys Acta. 1998 May 8;1380(3):377–388. doi: 10.1016/s0304-4165(98)00010-5. [DOI] [PubMed] [Google Scholar]
  2. Asplund T., Heldin P. Hyaluronan receptors are expressed on human malignant mesothelioma cells but not on normal mesothelial cells. Cancer Res. 1994 Aug 15;54(16):4516–4523. [PubMed] [Google Scholar]
  3. Asplund T., Versnel M. A., Laurent T. C., Heldin P. Human mesothelioma cells produce factors that stimulate the production of hyaluronan by mesothelial cells and fibroblasts. Cancer Res. 1993 Jan 15;53(2):388–392. [PubMed] [Google Scholar]
  4. Becq F., Jensen T. J., Chang X. B., Savoia A., Rommens J. M., Tsui L. C., Buchwald M., Riordan J. R., Hanrahan J. W. Phosphatase inhibitors activate normal and defective CFTR chloride channels. Proc Natl Acad Sci U S A. 1994 Sep 13;91(19):9160–9164. doi: 10.1073/pnas.91.19.9160. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Brinck J., Heldin P. Expression of recombinant hyaluronan synthase (HAS) isoforms in CHO cells reduces cell migration and cell surface CD44. Exp Cell Res. 1999 Nov 1;252(2):342–351. doi: 10.1006/excr.1999.4645. [DOI] [PubMed] [Google Scholar]
  6. DeAngelis P. L., Achyuthan A. M. Yeast-derived recombinant DG42 protein of Xenopus can synthesize hyaluronan in vitro. J Biol Chem. 1996 Sep 27;271(39):23657–23660. doi: 10.1074/jbc.271.39.23657. [DOI] [PubMed] [Google Scholar]
  7. DeAngelis P. L., Jing W., Drake R. R., Achyuthan A. M. Identification and molecular cloning of a unique hyaluronan synthase from Pasteurella multocida. J Biol Chem. 1998 Apr 3;273(14):8454–8458. doi: 10.1074/jbc.273.14.8454. [DOI] [PubMed] [Google Scholar]
  8. DeAngelis P. L., Jing W., Graves M. V., Burbank D. E., Van Etten J. L. Hyaluronan synthase of chlorella virus PBCV-1. Science. 1997 Dec 5;278(5344):1800–1803. doi: 10.1126/science.278.5344.1800. [DOI] [PubMed] [Google Scholar]
  9. DeAngelis P. L., Papaconstantinou J., Weigel P. H. Molecular cloning, identification, and sequence of the hyaluronan synthase gene from group A Streptococcus pyogenes. J Biol Chem. 1993 Sep 15;268(26):19181–19184. [PubMed] [Google Scholar]
  10. Heldin P., Asplund T., Ytterberg D., Thelin S., Laurent T. C. Characterization of the molecular mechanism involved in the activation of hyaluronan synthetase by platelet-derived growth factor in human mesothelial cells. Biochem J. 1992 Apr 1;283(Pt 1):165–170. doi: 10.1042/bj2830165. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Heldin P., Laurent T. C., Heldin C. H. Effect of growth factors on hyaluronan synthesis in cultured human fibroblasts. Biochem J. 1989 Mar 15;258(3):919–922. doi: 10.1042/bj2580919. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Honda A., Noguchi N., Takehara H., Ohashi Y., Asuwa N., Mori Y. Cooperative enhancement of hyaluronic acid synthesis by combined use of IGF-I and EGF, and inhibition by tyrosine kinase inhibitor genistein, in cultured mesothelial cells from rabbit pericardial cavity. J Cell Sci. 1991 Jan;98(Pt 1):91–98. doi: 10.1242/jcs.98.1.91. [DOI] [PubMed] [Google Scholar]
  13. Itano N., Kimata K. Expression cloning and molecular characterization of HAS protein, a eukaryotic hyaluronan synthase. J Biol Chem. 1996 Apr 26;271(17):9875–9878. doi: 10.1074/jbc.271.17.9875. [DOI] [PubMed] [Google Scholar]
  14. Klewes L., Prehm P. Intracellular signal transduction for serum activation of the hyaluronan synthase in eukaryotic cell lines. J Cell Physiol. 1994 Sep;160(3):539–544. doi: 10.1002/jcp.1041600317. [DOI] [PubMed] [Google Scholar]
  15. Knudson C. B., Knudson W. Hyaluronan-binding proteins in development, tissue homeostasis, and disease. FASEB J. 1993 Oct;7(13):1233–1241. [PubMed] [Google Scholar]
  16. Laurent T. C., Fraser J. R. Hyaluronan. FASEB J. 1992 Apr;6(7):2397–2404. [PubMed] [Google Scholar]
  17. Myers S. L., Brandt K. D. Effects of synovial fluid hyaluronan concentration and molecular size on clearance of protein from the canine knee. J Rheumatol. 1995 Sep;22(9):1732–1739. [PubMed] [Google Scholar]
  18. Roboz J., Greaves J., Silides D., Chahinian A. P., Holland J. F. Hyaluronic acid content of effusions as a diagnostic aid for malignant mesothelioma. Cancer Res. 1985 Apr;45(4):1850–1854. [PubMed] [Google Scholar]
  19. Rooney P., Kumar S., Ponting J., Wang M. The role of hyaluronan in tumour neovascularization (review). Int J Cancer. 1995 Mar 3;60(5):632–636. doi: 10.1002/ijc.2910600511. [DOI] [PubMed] [Google Scholar]
  20. Sherman L., Sleeman J., Herrlich P., Ponta H. Hyaluronate receptors: key players in growth, differentiation, migration and tumor progression. Curr Opin Cell Biol. 1994 Oct;6(5):726–733. doi: 10.1016/0955-0674(94)90100-7. [DOI] [PubMed] [Google Scholar]
  21. Shyjan A. M., Heldin P., Butcher E. C., Yoshino T., Briskin M. J. Functional cloning of the cDNA for a human hyaluronan synthase. J Biol Chem. 1996 Sep 20;271(38):23395–23399. doi: 10.1074/jbc.271.38.23395. [DOI] [PubMed] [Google Scholar]
  22. Spicer A. P., Augustine M. L., McDonald J. A. Molecular cloning and characterization of a putative mouse hyaluronan synthase. J Biol Chem. 1996 Sep 20;271(38):23400–23406. doi: 10.1074/jbc.271.38.23400. [DOI] [PubMed] [Google Scholar]
  23. Spicer A. P., McDonald J. A. Characterization and molecular evolution of a vertebrate hyaluronan synthase gene family. J Biol Chem. 1998 Jan 23;273(4):1923–1932. doi: 10.1074/jbc.273.4.1923. [DOI] [PubMed] [Google Scholar]
  24. Spicer A. P., Olson J. S., McDonald J. A. Molecular cloning and characterization of a cDNA encoding the third putative mammalian hyaluronan synthase. J Biol Chem. 1997 Apr 4;272(14):8957–8961. doi: 10.1074/jbc.272.14.8957. [DOI] [PubMed] [Google Scholar]
  25. Spicer A. P., Seldin M. F., Olsen A. S., Brown N., Wells D. E., Doggett N. A., Itano N., Kimata K., Inazawa J., McDonald J. A. Chromosomal localization of the human and mouse hyaluronan synthase genes. Genomics. 1997 May 1;41(3):493–497. doi: 10.1006/geno.1997.4696. [DOI] [PubMed] [Google Scholar]
  26. Sugiyama Y., Shimada A., Sayo T., Sakai S., Inoue S. Putative hyaluronan synthase mRNA are expressed in mouse skin and TGF-beta upregulates their expression in cultured human skin cells. J Invest Dermatol. 1998 Feb;110(2):116–121. doi: 10.1046/j.1523-1747.1998.00093.x. [DOI] [PubMed] [Google Scholar]
  27. Suzuki M., Asplund T., Yamashita H., Heldin C. H., Heldin P. Stimulation of hyaluronan biosynthesis by platelet-derived growth factor-BB and transforming growth factor-beta 1 involves activation of protein kinase C. Biochem J. 1995 May 1;307(Pt 3):817–821. doi: 10.1042/bj3070817. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Tengblad A. Quantitative analysis of hyaluronate in nanogram amounts. Biochem J. 1980 Jan 1;185(1):101–105. doi: 10.1042/bj1850101. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Tirone E., D'Alessandris C., Hascall V. C., Siracusa G., Salustri A. Hyaluronan synthesis by mouse cumulus cells is regulated by interactions between follicle-stimulating hormone (or epidermal growth factor) and a soluble oocyte factor (or transforming growth factor beta1). J Biol Chem. 1997 Feb 21;272(8):4787–4794. doi: 10.1074/jbc.272.8.4787. [DOI] [PubMed] [Google Scholar]
  30. Toole B. P. Hyaluronan and its binding proteins, the hyaladherins. Curr Opin Cell Biol. 1990 Oct;2(5):839–844. doi: 10.1016/0955-0674(90)90081-o. [DOI] [PubMed] [Google Scholar]
  31. Watanabe K., Yamaguchi Y. Molecular identification of a putative human hyaluronan synthase. J Biol Chem. 1996 Sep 20;271(38):22945–22948. doi: 10.1074/jbc.271.38.22945. [DOI] [PubMed] [Google Scholar]
  32. Weigel P. H., Hascall V. C., Tammi M. Hyaluronan synthases. J Biol Chem. 1997 May 30;272(22):13997–14000. doi: 10.1074/jbc.272.22.13997. [DOI] [PubMed] [Google Scholar]
  33. Wells A. F., Klareskog L., Lindblad S., Laurent T. C. Correlation between increased hyaluronan localized in arthritic synovium and the presence of proliferating cells. A role for macrophage-derived factors. Arthritis Rheum. 1992 Apr;35(4):391–396. doi: 10.1002/art.1780350405. [DOI] [PubMed] [Google Scholar]
  34. West D. C., Hampson I. N., Arnold F., Kumar S. Angiogenesis induced by degradation products of hyaluronic acid. Science. 1985 Jun 14;228(4705):1324–1326. doi: 10.1126/science.2408340. [DOI] [PubMed] [Google Scholar]
  35. Yamada Y., Itano N., Zako M., Yoshida M., Lenas P., Niimi A., Ueda M., Kimata K. The gene structure and promoter sequence of mouse hyaluronan synthase 1. Biochem J. 1998 Mar 15;330(Pt 3):1223–1227. doi: 10.1042/bj3301223. [DOI] [PMC free article] [PubMed] [Google Scholar]

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