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. 1992 Oct 15;89(20):9691–9694. doi: 10.1073/pnas.89.20.9691

Direct stimulation of immediate-early genes by intranuclear insulin in trypsin-treated H35 hepatoma cells.

Y J Lin 1, S Harada 1, E G Loten 1, R M Smith 1, L Jarett 1
PMCID: PMC50198  PMID: 1409684

Abstract

H35 hepatoma cells were treated with trypsin to abolish insulin binding and insulin-stimulated receptor kinase activity. Insulin was, however, internalized by fluid-phase endocytosis in trypsin-treated cells. Furthermore, nuclear accumulation of insulin was similar in control and trypsin-treated hepatoma cells. Northern blot analysis revealed insulin increased g33 and c-fos mRNA concentrations identically in control and trypsin-treated cells but had no effect on beta 2-microglobulin mRNA. Actinomycin D treatment prior to or after insulin addition demonstrated that insulin increased gene transcription and had no effect on mRNA degradation. These studies suggest that the accumulation of intact insulin in cell nuclei may be directly involved in the increased transcription of immediate-early genes.

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Selected References

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  1. Cao H., Decker S., Stern D. F. TPA inhibits the tyrosine kinase activity of the neu protein in vivo and in vitro. Oncogene. 1991 May;6(5):705–711. [PubMed] [Google Scholar]
  2. Chomczynski P., Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987 Apr;162(1):156–159. doi: 10.1006/abio.1987.9999. [DOI] [PubMed] [Google Scholar]
  3. Chu D. T., Davis C. M., Chrapkiewicz N. B., Granner D. K. Reciprocal regulation of gene transcription by insulin. Inhibition of the phosphoenolpyruvate carboxykinase gene and stimulation of gene 33 in a single cell type. J Biol Chem. 1988 Sep 15;263(26):13007–13011. [PubMed] [Google Scholar]
  4. Clark S., Larkins R. G., De Luise M., Melick R. A. The effects of trypsin and phospholipase C on insulin binding and action in the isolated adipocyte. Biochem J. 1980 Feb 15;186(2):535–540. doi: 10.1042/bj1860535. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Gottschalk W. K. The pathway mediating insulin's effects on pyruvate dehydrogenase bypasses the insulin receptor tyrosine kinase. J Biol Chem. 1991 May 15;266(14):8814–8819. [PubMed] [Google Scholar]
  6. Kasuga M., Karlsson F. A., Kahn C. R. Insulin stimulates the phosphorylation of the 95,000-dalton subunit of its own receptor. Science. 1982 Jan 8;215(4529):185–187. doi: 10.1126/science.7031900. [DOI] [PubMed] [Google Scholar]
  7. Kono T., Barham F. W. The relationship between the insulin-binding capacity of fat cells and the cellular response to insulin. Studies with intact and trypsin-treated fat cells. J Biol Chem. 1971 Oct 25;246(20):6210–6216. [PubMed] [Google Scholar]
  8. Lee K. L., Isham K. R., Stringfellow L., Rothrock R., Kenney F. T. Molecular cloning of cDNAs cognate to genes sensitive to hormonal control in rat liver. J Biol Chem. 1985 Dec 25;260(30):16433–16438. [PubMed] [Google Scholar]
  9. Messina J. L., Hamlin J., Larner J. Effects of insulin alone on the accumulation of a specific mRNA in rat hepatoma cells. J Biol Chem. 1985 Dec 25;260(30):16418–16423. [PubMed] [Google Scholar]
  10. Messina J. L. Insulin and dexamethasone regulation of a rat hepatoma messenger ribonucleic acid: insulin has a transcriptional and a posttranscriptional effect. Endocrinology. 1989 Feb;124(2):754–761. doi: 10.1210/endo-124-2-754. [DOI] [PubMed] [Google Scholar]
  11. Messina J. L. Insulin's regulation of c-fos gene transcription in hepatoma cells. J Biol Chem. 1990 Jul 15;265(20):11700–11705. [PubMed] [Google Scholar]
  12. Miller D. S. Stimulation of RNA and protein synthesis by intracellular insulin. Science. 1988 Apr 22;240(4851):506–509. doi: 10.1126/science.2451860. [DOI] [PubMed] [Google Scholar]
  13. Mohn K. L., Laz T. M., Melby A. E., Taub R. Immediate-early gene expression differs between regenerating liver, insulin-stimulated H-35 cells, and mitogen-stimulated Balb/c 3T3 cells. Liver-specific induction patterns of gene 33, phosphoenolpyruvate carboxykinase, and the jun, fos, and egr families. J Biol Chem. 1990 Dec 15;265(35):21914–21921. [PubMed] [Google Scholar]
  14. Moller D. E., Benecke H., Flier J. S. Biologic activities of naturally occurring human insulin receptor mutations. Evidence that metabolic effects of insulin can be mediated by a kinase-deficient insulin receptor mutant. J Biol Chem. 1991 Jun 15;266(17):10995–11001. [PubMed] [Google Scholar]
  15. Moss A. L., Ward W. F. Multiple pathways for ligand internalization in rat hepatocytes. I: Effects of anoxia, phenylarsine oxide and monensin. J Cell Physiol. 1991 Nov;149(2):313–318. doi: 10.1002/jcp.1041490219. [DOI] [PubMed] [Google Scholar]
  16. Moss A. L., Ward W. F. Multiple pathways for ligand internalization in rat hepatocytes. II: Effect of hyperosmolarity and contribution of fluid-phase endocytosis. J Cell Physiol. 1991 Nov;149(2):319–323. doi: 10.1002/jcp.1041490220. [DOI] [PubMed] [Google Scholar]
  17. Murakami M. S., Rosen O. M. The role of insulin receptor autophosphorylation in signal transduction. J Biol Chem. 1991 Nov 25;266(33):22653–22660. [PubMed] [Google Scholar]
  18. O'Brien R. M., Granner D. K. Regulation of gene expression by insulin. Biochem J. 1991 Sep 15;278(Pt 3):609–619. doi: 10.1042/bj2780609. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Olefsky J. M. The insulin receptor. A multifunctional protein. Diabetes. 1990 Sep;39(9):1009–1016. doi: 10.2337/diab.39.9.1009. [DOI] [PubMed] [Google Scholar]
  20. Pilch P. F., Axelrod J. D., Colello J., Czech M. P. Unimpaired signal transduction by the adipocyte insulin receptor following its partial proteolytic fragmentation. J Biol Chem. 1981 Feb 25;256(4):1570–1575. [PubMed] [Google Scholar]
  21. Purrello F., Vigneri R., Clawson G. A., Goldfine I. D. Insulin stimulation of nucleoside triphosphatase activity in isolated nuclear envelopes. Science. 1982 May 28;216(4549):1005–1007. doi: 10.1126/science.6281885. [DOI] [PubMed] [Google Scholar]
  22. Rosen O. M. After insulin binds. Science. 1987 Sep 18;237(4821):1452–1458. doi: 10.1126/science.2442814. [DOI] [PubMed] [Google Scholar]
  23. Schumm D. E., Webb T. E. Effect of physiological concentrations of insulin and antidiabetic drugs on RNA release from isolated liver nuclei. J Cell Biochem. 1983;23(1-4):223–229. doi: 10.1002/jcb.240230119. [DOI] [PubMed] [Google Scholar]
  24. Shoelson S. E., White M. F., Kahn C. R. Tryptic activation of the insulin receptor. Proteolytic truncation of the alpha-subunit releases the beta-subunit from inhibitory control. J Biol Chem. 1988 Apr 5;263(10):4852–4860. [PubMed] [Google Scholar]
  25. Smith R. M., Jarett L. Partial characterization of mechanism of insulin accumulation in H35 hepatoma cell nuclei. Diabetes. 1990 Jun;39(6):683–689. doi: 10.2337/diab.39.6.683. [DOI] [PubMed] [Google Scholar]
  26. Smith R. M., Jarett L. Ultrastructural evidence for the accumulation of insulin in nuclei of intact 3T3-L1 adipocytes by an insulin-receptor mediated process. Proc Natl Acad Sci U S A. 1987 Jan;84(2):459–463. doi: 10.1073/pnas.84.2.459. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Soler A. P., Alemany J., Smith R. M., de Pablo F., Jarett L. The state of differentiation of embryonic chicken lens cells determines insulin-like growth factor I internalization. Endocrinology. 1990 Aug;127(2):595–603. doi: 10.1210/endo-127-2-595. [DOI] [PubMed] [Google Scholar]
  28. Soler A. P., Smith R. M., Jarett L. Insulin stimulates accumulation and efflux of macromolecules in isolated nuclei from H35 hepatoma cells. Diabetes. 1992 Feb;41(2):194–201. doi: 10.2337/diab.41.2.194. [DOI] [PubMed] [Google Scholar]
  29. Soler A. P., Thompson K. A., Smith R. M., Jarett L. Immunological demonstration of the accumulation of insulin, but not insulin receptors, in nuclei of insulin-treated cells. Proc Natl Acad Sci U S A. 1989 Sep;86(17):6640–6644. doi: 10.1073/pnas.86.17.6640. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Taub R., Roy A., Dieter R., Koontz J. Insulin as a growth factor in rat hepatoma cells. Stimulation of proto-oncogene expression. J Biol Chem. 1987 Aug 5;262(22):10893–10897. [PubMed] [Google Scholar]
  31. Thompson K. A., Soler A. P., Smith R. M., Jarett L. Intranuclear localization of insulin in rat hepatoma cells: insulin/matrix association. Eur J Cell Biol. 1989 Dec;50(2):442–446. [PubMed] [Google Scholar]
  32. Zhang B., Tavaré J. M., Ellis L., Roth R. A. The regulatory role of known tyrosine autophosphorylation sites of the insulin receptor kinase domain. An assessment by replacement with neutral and negatively charged amino acids. J Biol Chem. 1991 Jan 15;266(2):990–996. [PubMed] [Google Scholar]
  33. de Vries C. P., Van der Veen E. A. Characteristics of insulin binding to H35 hepatoma cells. Biochimie. 1985 Oct-Nov;67(10-11):1191–1194. doi: 10.1016/s0300-9084(85)80119-x. [DOI] [PubMed] [Google Scholar]

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