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. 1987 Jan;84(1):126–130. doi: 10.1073/pnas.84.1.126

Homologous down-regulation of the insulin receptor is associated with increased receptor biosynthesis in cultured human lymphocytes (IM-9 line).

D G Rouiller, P Gorden
PMCID: PMC304155  PMID: 3099291

Abstract

Cultured IM-9 lymphocytes were preincubated with 1 microM insulin, a condition resulting in a 56% reduction in cell surface insulin receptors. Cellular proteins were then metabolically labeled, and the radioactivity incorporated into the insulin proreceptor and receptor mature subunits was measured over a 4-hr chase period. As early as 30 min of chase, incorporation into the proreceptor was 28 +/- 6% higher in down-regulated cells than in control cells (mean +/- SEM, P less than 0.05). By 1 hr of chase, the difference reached 41 +/- 14% for the proreceptor and 84 +/- 28% for the alpha subunit (P less than 0.01); values returned to normal by 2 hr. At 4 hr of chase, labeling of the alpha subunit of down-regulated cells was diminished 36 +/- 9% below control (P less than 0.05). The increased biosynthetic rate of the proreceptor was more prominent when the chase medium contained 25 microM monensin, an inhibitor of processing of the proreceptor into mature subunits. Similar effects occurred whether [3H]mannose or [3H]lysine was used as biosynthetic marker. The effect was specific for the insulin receptor. These data demonstrate that insulin receptor homologous down-regulation is associated with increased proreceptor biosynthesis and processing into mature subunits. This might represent a cellular mechanism compensating for insulin-induced receptor loss.

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Selected References

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

  1. Carpentier J. L., Dayer J. M., Lang U., Silverman R., Orci L., Gorden P. Down-regulation and recycling of insulin receptors. Effect of monensin on IM-9 lymphocytes and U-937 monocyte-like cells. J Biol Chem. 1984 Nov 25;259(22):14190–14195. [PubMed] [Google Scholar]
  2. Clark A. J., Ishii S., Richert N., Merlino G. T., Pastan I. Epidermal growth factor regulates the expression of its own receptor. Proc Natl Acad Sci U S A. 1985 Dec;82(24):8374–8378. doi: 10.1073/pnas.82.24.8374. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Ebina Y., Ellis L., Jarnagin K., Edery M., Graf L., Clauser E., Ou J. H., Masiarz F., Kan Y. W., Goldfine I. D. The human insulin receptor cDNA: the structural basis for hormone-activated transmembrane signalling. Cell. 1985 Apr;40(4):747–758. doi: 10.1016/0092-8674(85)90334-4. [DOI] [PubMed] [Google Scholar]
  4. Fehlmann M., Carpentier J. L., Van Obberghen E., Freychet P., Thamm P., Saunders D., Brandenburg D., Orci L. Internalized insulin receptors are recycled to the cell surface in rat hepatocytes. Proc Natl Acad Sci U S A. 1982 Oct;79(19):5921–5925. doi: 10.1073/pnas.79.19.5921. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Gavin J. R., 3rd, Roth J., Neville D. M., Jr, de Meyts P., Buell D. N. Insulin-dependent regulation of insulin receptor concentrations: a direct demonstration in cell culture. Proc Natl Acad Sci U S A. 1974 Jan;71(1):84–88. doi: 10.1073/pnas.71.1.84. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Goldstein J. L., Brown M. S., Anderson R. G., Russell D. W., Schneider W. J. Receptor-mediated endocytosis: concepts emerging from the LDL receptor system. Annu Rev Cell Biol. 1985;1:1–39. doi: 10.1146/annurev.cb.01.110185.000245. [DOI] [PubMed] [Google Scholar]
  7. Gorden P., Carpentier J. L., Freychet P. O., Orci L. Internalization of polypeptide hormones: mechanism, intracellular localization and significance. Diabetologia. 1980 Apr;18(4):263–274. doi: 10.1007/BF00251003. [DOI] [PubMed] [Google Scholar]
  8. Gorden P., Carpentier J. L., Orci L. Insulin action at the cellular level: anatomical considerations. Diabetes Metab Rev. 1985;1(1-2):99–117. doi: 10.1002/dmr.5610010106. [DOI] [PubMed] [Google Scholar]
  9. Harrison L. C., Flier J. S., Roth J., Karlsson F. A., Kahn C. R. Immunoprecipitation of the insulin receptor: a sensitive assay for receptor antibodies and a specific technique for receptor purification. J Clin Endocrinol Metab. 1979 Jan;48(1):59–65. doi: 10.1210/jcem-48-1-59. [DOI] [PubMed] [Google Scholar]
  10. Hedo J. A., Gorden P. Biosynthesis of the insulin receptor. Horm Metab Res. 1985 Oct;17(10):487–490. doi: 10.1055/s-2007-1013586. [DOI] [PubMed] [Google Scholar]
  11. Hedo J. A., Kahn C. R., Hayashi M., Yamada K. M., Kasuga M. Biosynthesis and glycosylation of the insulin receptor. Evidence for a single polypeptide precursor of the two major subunits. J Biol Chem. 1983 Aug 25;258(16):10020–10026. [PubMed] [Google Scholar]
  12. Jacobs S., Kull F. C., Jr, Cuatrecasas P. Monensin blocks the maturation of receptors for insulin and somatomedin C: identification of receptor precursors. Proc Natl Acad Sci U S A. 1983 Mar;80(5):1228–1231. doi: 10.1073/pnas.80.5.1228. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Kasuga M., Kahn C. R., Hedo J. A., Van Obberghen E., Yamada K. M. Insulin-induced receptor loss in cultured human lymphocytes is due to accelerated receptor degradation. Proc Natl Acad Sci U S A. 1981 Nov;78(11):6917–6921. doi: 10.1073/pnas.78.11.6917. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Knutson V. P., Ronnett G. V., Lane M. D. Control of insulin receptor level in 3T3 cells: effect of insulin-induced down-regulation and dexamethasone-induced up-regulation on rate of receptor inactivation. Proc Natl Acad Sci U S A. 1982 May;79(9):2822–2826. doi: 10.1073/pnas.79.9.2822. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Krupp M. N., Connolly D. T., Lane M. D. Synthesis, turnover, and down-regulation of epidermal growth factor receptors in human A431 epidermoid carcinoma cells and skin fibroblasts. J Biol Chem. 1982 Oct 10;257(19):11489–11496. [PubMed] [Google Scholar]
  16. Krupp M., Lane M. D. On the mechanism of ligand-induced down-regulation of insulin receptor level in the liver cell. J Biol Chem. 1981 Feb 25;256(4):1689–1694. [PubMed] [Google Scholar]
  17. Kudlow J. E., Cheung C. Y., Bjorge J. D. Epidermal growth factor stimulates the synthesis of its own receptor in a human breast cancer cell line. J Biol Chem. 1986 Mar 25;261(9):4134–4138. [PubMed] [Google Scholar]
  18. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  19. Ronnett G. V., Knutson V. P., Lane M. D. Insulin-induced down-regulation of insulin receptors in 3T3-L1 adipocytes. Altered rate of receptor inactivation. J Biol Chem. 1982 Apr 25;257(8):4285–4291. [PubMed] [Google Scholar]
  20. Rouiller D. G., McElduff A., Hedo J. A., Gorden P. Induction of the insulin proreceptor by hydrocortisone in cultured lymphocytes (IM-9 line). J Clin Invest. 1985 Aug;76(2):645–649. doi: 10.1172/JCI112017. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Rubin C. S., Hirsch A., Fung C., Rosen O. M. Development of hormone receptors and hormonal responsiveness in vitro. Insulin receptors and insulin sensitivity in the preadipocyte and adipocyte forms of 3T3-L1 cells. J Biol Chem. 1978 Oct 25;253(20):7570–7578. [PubMed] [Google Scholar]
  22. Russell D. W., Yamamoto T., Schneider W. J., Slaughter C. J., Brown M. S., Goldstein J. L. cDNA cloning of the bovine low density lipoprotein receptor: feedback regulation of a receptor mRNA. Proc Natl Acad Sci U S A. 1983 Dec;80(24):7501–7505. doi: 10.1073/pnas.80.24.7501. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Ueda M., Robinson F. W., Smith M. M., Kono T. Effects of monensin on insulin processing in adipocytes. Evidence that the internalized insulin-receptor complex has some physiological activities. J Biol Chem. 1985 Apr 10;260(7):3941–3946. [PubMed] [Google Scholar]
  24. Ullrich A., Bell J. R., Chen E. Y., Herrera R., Petruzzelli L. M., Dull T. J., Gray A., Coussens L., Liao Y. C., Tsubokawa M. Human insulin receptor and its relationship to the tyrosine kinase family of oncogenes. 1985 Feb 28-Mar 6Nature. 313(6005):756–761. doi: 10.1038/313756a0. [DOI] [PubMed] [Google Scholar]

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