Skip to main content
NCBI home page
Biochemical Journal logoLink to Biochemical Journal
. 1989 Dec 15;264(3):813–822. doi: 10.1042/bj2640813

Receptor-mediated endocytosis and nuclear transport of human interleukin 1 alpha.

S Grenfell 1, N Smithers 1, K Miller 1, R Solari 1
PMCID: PMC1133658  PMID: 2533500

Abstract

In this study we demonstrate that 125I-labelled interleukin (IL) 1 alpha binds specifically to its receptor on the surface of EL4 6.1 cells and is subsequently endocytosed and translocated from the cell membrane to the nucleus, where it progressively accumulates. Two-dimensional polyacrylamide-gel electrophoresis revealed that the internalized 125I-IL1 alpha associated with the nucleus was intact, with negligible breakdown products present. Specific and saturable binding of 125I-IL1 alpha was demonstrated on purified nuclei isolated from these cells. Binding of the radiolabelled ligand showed similar kinetics to that of the plasma-membrane receptor, and was inhibited by both unlabelled IL1 alpha and IL1 beta. Equilibrium binding studies on isolated nuclei revealed a single high-affinity binding site, with a Kd of 17 +/- 2 pM, and 79 +/- 12 binding sites per nucleus. These studies demonstrate that receptor-mediated endocytosis of IL1 results in its accumulation in the nucleus, and this mechanism may play an important role in mediating some of the actions of IL1.

Full text

PDF
813

Images in this article

816Fig. 4.
on p.816
818Fig. 5.
on p.818

Selected References

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

  1. Abraham R. T., Ho S. N., Barna T. J., McKean D. J. Transmembrane signaling during interleukin 1-dependent T cell activation. Interactions of signal 1- and signal 2-type mediators with the phosphoinositide-dependent signal transduction mechanism. J Biol Chem. 1987 Feb 25;262(6):2719–2728. [PubMed] [Google Scholar]
  2. Auron P. E., Webb A. C., Rosenwasser L. J., Mucci S. F., Rich A., Wolff S. M., Dinarello C. A. Nucleotide sequence of human monocyte interleukin 1 precursor cDNA. Proc Natl Acad Sci U S A. 1984 Dec;81(24):7907–7911. doi: 10.1073/pnas.81.24.7907. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bird T. A., Saklatvala J. Studies on the fate of receptor-bound 125I-interleukin 1 beta in porcine synovial fibroblasts. J Immunol. 1987 Jul 1;139(1):92–97. [PubMed] [Google Scholar]
  4. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
  5. Carpentier J. L., Gazzano H., Van Obberghen E., Fehlmann M., Freychet P., Orci L. Intracellular pathway followed by the insulin receptor covalently coupled to 125I-photoreactive insulin during internalization and recycling. J Cell Biol. 1986 Mar;102(3):989–996. doi: 10.1083/jcb.102.3.989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Carroll G. J. Porcine catabolin stimulates prostaglandin E2 secretion but does not affect intracellular cyclic AMP production in pig synovial fibroblasts. Ann Rheum Dis. 1985 Sep;44(9):631–636. doi: 10.1136/ard.44.9.631. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Chedid M., Shirakawa F., Naylor P., Mizel S. B. Signal transduction pathway for IL-1. Involvement of a pertussis toxin-sensitive GTP-binding protein in the activation of adenylate cyclase. J Immunol. 1989 Jun 15;142(12):4301–4306. [PubMed] [Google Scholar]
  8. Colotta F., Lampugnani M. G., Polentarutti N., Dejana E., Mantovani A. Interleukin-1 induces c-fos protooncogene expression in cultured human endothelial cells. Biochem Biophys Res Commun. 1988 May 16;152(3):1104–1110. doi: 10.1016/s0006-291x(88)80398-x. [DOI] [PubMed] [Google Scholar]
  9. Davis C. G., Goldstein J. L., Südhof T. C., Anderson R. G., Russell D. W., Brown M. S. Acid-dependent ligand dissociation and recycling of LDL receptor mediated by growth factor homology region. Nature. 1987 Apr 23;326(6115):760–765. doi: 10.1038/326760a0. [DOI] [PubMed] [Google Scholar]
  10. Dinarello C. A. Biology of interleukin 1. FASEB J. 1988 Feb;2(2):108–115. [PubMed] [Google Scholar]
  11. Dingwall C., Laskey R. A. Protein import into the cell nucleus. Annu Rev Cell Biol. 1986;2:367–390. doi: 10.1146/annurev.cb.02.110186.002055. [DOI] [PubMed] [Google Scholar]
  12. Dingwall C., Robbins J., Dilworth S. M., Roberts B., Richardson W. D. The nucleoplasmin nuclear location sequence is larger and more complex than that of SV-40 large T antigen. J Cell Biol. 1988 Sep;107(3):841–849. doi: 10.1083/jcb.107.3.841. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Dunn W. A., Connolly T. P., Hubbard A. L. Receptor-mediated endocytosis of epidermal growth factor by rat hepatocytes: receptor pathway. J Cell Biol. 1986 Jan;102(1):24–36. doi: 10.1083/jcb.102.1.24. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Dunn W. A., Hubbard A. L. Receptor-mediated endocytosis of epidermal growth factor by hepatocytes in the perfused rat liver: ligand and receptor dynamics. J Cell Biol. 1984 Jun;98(6):2148–2159. doi: 10.1083/jcb.98.6.2148. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Duprez V., Cornet V., Dautry-Varsat A. Down-regulation of high affinity interleukin 2 receptors in a human tumor T cell line. Interleukin 2 increases the rate of surface receptor decay. J Biol Chem. 1988 Sep 15;263(26):12860–12865. [PubMed] [Google Scholar]
  16. Frisch S. M., Ruley H. E. Transcription from the stromelysin promoter is induced by interleukin-1 and repressed by dexamethasone. J Biol Chem. 1987 Dec 5;262(34):16300–16304. [PubMed] [Google Scholar]
  17. Georgilis K., Schaefer C., Dinarello C. A., Klempner M. S. Human recombinant interleukin 1 beta has no effect on intracellular calcium or on functional responses of human neutrophils. J Immunol. 1987 May 15;138(10):3403–3407. [PubMed] [Google Scholar]
  18. Goldfine I. D., Smith G. J., Wong K. Y., Jones A. L. Cellular uptake and nuclear binding of insulin in human cultured lymphocytes: evidence for potential intracellular sites of insulin action. Proc Natl Acad Sci U S A. 1977 Apr;74(4):1368–1372. doi: 10.1073/pnas.74.4.1368. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. 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]
  20. Hagiwara H., Huang H. J., Arai N., Herzenberg L. A., Arai K., Zlotnik A. Interleukin 1 modulates messenger RNA levels of lymphokines and of other molecules associated with T cell activation in the T cell lymphoma LBRM33-1A5. J Immunol. 1987 Apr 15;138(8):2514–2519. [PubMed] [Google Scholar]
  21. Hanover J. A., Willingham M. C., Pastan I. Kinetics of transit of transferrin and epidermal growth factor through clathrin-coated membranes. Cell. 1984 Dec;39(2 Pt 1):283–293. doi: 10.1016/0092-8674(84)90006-0. [DOI] [PubMed] [Google Scholar]
  22. Heino J., Kähäri V. M., Mauviel A., Krusius T. Human recombinant interleukin-1 regulates cellular mRNA levels of dermatan sulphate proteoglycan core protein. Biochem J. 1988 May 15;252(1):309–312. doi: 10.1042/bj2520309. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Hopkins C. R. Intracellular routing of transferrin and transferrin receptors in epidermoid carcinoma A431 cells. Cell. 1983 Nov;35(1):321–330. doi: 10.1016/0092-8674(83)90235-0. [DOI] [PubMed] [Google Scholar]
  24. Hopkins C. R., Trowbridge I. S. Internalization and processing of transferrin and the transferrin receptor in human carcinoma A431 cells. J Cell Biol. 1983 Aug;97(2):508–521. doi: 10.1083/jcb.97.2.508. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Hubbard A. L., Ma A. Isolation of rat hepatocyte plasma membranes. II. Identification of membrane-associated cytoskeletal proteins. J Cell Biol. 1983 Jan;96(1):230–239. doi: 10.1083/jcb.96.1.230. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Johnson L. K., Vlodavsky I., Baxter J. D., Gospodarowicz D. Nuclear accumulation of epidermal growth factor in cultured rat pituitary cells. Nature. 1980 Sep 25;287(5780):340–343. doi: 10.1038/287340a0. [DOI] [PubMed] [Google Scholar]
  27. Kester M., Simonson M. S., Mené P., Sedor J. R. Interleukin-1 generates transmembrane signals from phospholipids through novel pathways in cultured rat mesangial cells. J Clin Invest. 1989 Feb;83(2):718–723. doi: 10.1172/JCI113937. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Kilian P. L., Kaffka K. L., Stern A. S., Woehle D., Benjamin W. R., Dechiara T. M., Gubler U., Farrar J. J., Mizel S. B., Lomedico P. T. Interleukin 1 alpha and interleukin 1 beta bind to the same receptor on T cells. J Immunol. 1986 Jun 15;136(12):4509–4514. [PubMed] [Google Scholar]
  29. Kovacs E. J., Oppenheim J. J., Young H. A. Induction of c-fos and c-myc expression in T lymphocytes after treatment with recombinant interleukin 1-alpha. J Immunol. 1986 Dec 1;137(11):3649–3651. [PubMed] [Google Scholar]
  30. Lin J. X., Vilcek J. Tumor necrosis factor and interleukin-1 cause a rapid and transient stimulation of c-fos and c-myc mRNA levels in human fibroblasts. J Biol Chem. 1987 Sep 5;262(25):11908–11911. [PubMed] [Google Scholar]
  31. Lomedico P. T., Gubler U., Hellmann C. P., Dukovich M., Giri J. G., Pan Y. C., Collier K., Semionow R., Chua A. O., Mizel S. B. Cloning and expression of murine interleukin-1 cDNA in Escherichia coli. 1984 Nov 29-Dec 5Nature. 312(5993):458–462. doi: 10.1038/312458a0. [DOI] [PubMed] [Google Scholar]
  32. Lomedico P. T., Kilian P. L., Gubler U., Stern A. S., Chizzonite R. Molecular biology of interleukin-1. Cold Spring Harb Symp Quant Biol. 1986;51(Pt 1):631–639. doi: 10.1101/sqb.1986.051.01.075. [DOI] [PubMed] [Google Scholar]
  33. Lowenthal J. W., MacDonald H. R. Binding and internalization of interleukin 1 by T cells. Direct evidence for high- and low-affinity classes of interleukin 1 receptor. J Exp Med. 1986 Oct 1;164(4):1060–1074. doi: 10.1084/jem.164.4.1060. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. March C. J., Mosley B., Larsen A., Cerretti D. P., Braedt G., Price V., Gillis S., Henney C. S., Kronheim S. R., Grabstein K. Cloning, sequence and expression of two distinct human interleukin-1 complementary DNAs. Nature. 1985 Jun 20;315(6021):641–647. doi: 10.1038/315641a0. [DOI] [PubMed] [Google Scholar]
  35. Martin M., Resch K. Interleukin 1: more than a mediator between leukocytes. Trends Pharmacol Sci. 1988 May;9(5):171–177. doi: 10.1016/0165-6147(88)90033-8. [DOI] [PubMed] [Google Scholar]
  36. Mates G., Daniel M., Walker C. Factors affecting the reproducibility of a spectrofluorimetric assay for the enumeration of human venous endothelium in culture. Cell Biol Int Rep. 1986 Aug;10(8):641–648. doi: 10.1016/0309-1651(86)90141-4. [DOI] [PubMed] [Google Scholar]
  37. Matsushima K., Kobayashi Y., Copeland T. D., Akahoshi T., Oppenheim J. J. Phosphorylation of a cytosolic 65-kDa protein induced by interleukin 1 in glucocorticoid pretreated normal human peripheral blood mononuclear leukocytes. J Immunol. 1987 Nov 15;139(10):3367–3374. [PubMed] [Google Scholar]
  38. Mauviel A., Teyton L., Bhatnagar R., Penfornis H., Laurent M., Hartmann D., Bonaventure J., Loyau G., Saklatvala J., Pujol J. P. Interleukin-1 alpha modulates collagen gene expression in cultured synovial cells. Biochem J. 1988 May 15;252(1):247–255. doi: 10.1042/bj2520247. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Mizel S. B., Kilian P. L., Lewis J. C., Paganelli K. A., Chizzonite R. A. The interleukin 1 receptor. Dynamics of interleukin 1 binding and internalization in T cells and fibroblasts. J Immunol. 1987 May 1;138(9):2906–2912. [PubMed] [Google Scholar]
  40. Nilsson J., Thyberg J., Heldin C. H., Westermark B., Wasteson A. Surface binding and internalization of platelet-derived growth factor in human fibroblasts. Proc Natl Acad Sci U S A. 1983 Sep;80(18):5592–5596. doi: 10.1073/pnas.80.18.5592. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. O'Farrell P. H. High resolution two-dimensional electrophoresis of proteins. J Biol Chem. 1975 May 25;250(10):4007–4021. [PMC free article] [PubMed] [Google Scholar]
  42. Perlmutter D. H., Goldberger G., Dinarello C. A., Mizel S. B., Colten H. R. Regulation of class III major histocompatibility complex gene products by interleukin-1. Science. 1986 May 16;232(4752):850–852. doi: 10.1126/science.3010455. [DOI] [PubMed] [Google Scholar]
  43. Podlecki D. A., Smith R. M., Kao M., Tsai P., Huecksteadt T., Brandenburg D., Lasher R. S., Jarett L., Olefsky J. M. Nuclear translocation of the insulin receptor. A possible mediator of insulin's long term effects. J Biol Chem. 1987 Mar 5;262(7):3362–3368. [PubMed] [Google Scholar]
  44. Qwarnstrom E. E., Page R. C., Gillis S., Dower S. K. Binding, internalization, and intracellular localization of interleukin-1 beta in human diploid fibroblasts. J Biol Chem. 1988 Jun 15;263(17):8261–8269. [PubMed] [Google Scholar]
  45. REYNOLDS E. S. The use of lead citrate at high pH as an electron-opaque stain in electron microscopy. J Cell Biol. 1963 Apr;17:208–212. doi: 10.1083/jcb.17.1.208. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Rakowicz-Szulczynska E. M., Rodeck U., Herlyn M., Koprowski H. Chromatin binding of epidermal growth factor, nerve growth factor, and platelet-derived growth factor in cells bearing the appropriate surface receptors. Proc Natl Acad Sci U S A. 1986 Jun;83(11):3728–3732. doi: 10.1073/pnas.83.11.3728. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Reed B. C., Glasted K., Miller B. Direct comparison of the rates of internalization and degradation of covalent receptor-insulin complexes in 3T3-L1 adipocytes. Internalization of occupied receptors is not the rate-limiting step in receptor-hormone complex degradation. J Biol Chem. 1984 Jul 10;259(13):8134–8143. [PubMed] [Google Scholar]
  48. Rosoff P. M., Savage N., Dinarello C. A. Interleukin-1 stimulates diacylglycerol production in T lymphocytes by a novel mechanism. Cell. 1988 Jul 1;54(1):73–81. doi: 10.1016/0092-8674(88)90181-x. [DOI] [PubMed] [Google Scholar]
  49. Salacinski P. R., McLean C., Sykes J. E., Clement-Jones V. V., Lowry P. J. Iodination of proteins, glycoproteins, and peptides using a solid-phase oxidizing agent, 1,3,4,6-tetrachloro-3 alpha,6 alpha-diphenyl glycoluril (Iodogen). Anal Biochem. 1981 Oct;117(1):136–146. doi: 10.1016/0003-2697(81)90703-x. [DOI] [PubMed] [Google Scholar]
  50. Shirakawa F., Yamashita U., Chedid M., Mizel S. B. Cyclic AMP--an intracellular second messenger for interleukin 1. Proc Natl Acad Sci U S A. 1988 Nov;85(21):8201–8205. doi: 10.1073/pnas.85.21.8201. [DOI] [PMC free article] [PubMed] [Google Scholar]
  51. Sims J. E., March C. J., Cosman D., Widmer M. B., MacDonald H. R., McMahan C. J., Grubin C. E., Wignall J. M., Jackson J. L., Call S. M. cDNA expression cloning of the IL-1 receptor, a member of the immunoglobulin superfamily. Science. 1988 Jul 29;241(4865):585–589. doi: 10.1126/science.2969618. [DOI] [PubMed] [Google Scholar]
  52. Tata J. R. Isolation of nuclei from liver and other tissues. Methods Enzymol. 1974;31:253–262. doi: 10.1016/0076-6879(74)31027-0. [DOI] [PubMed] [Google Scholar]
  53. Vale R. D., Ignatius M. J., Shooter E. M. Association of nerve growth factor receptors with the triton X-100 cytoskeleton of PC12 cells. J Neurosci. 1985 Oct;5(10):2762–2770. doi: 10.1523/JNEUROSCI.05-10-02762.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  54. Von Hoegen I., Falk W., Kojouharoff G., Krammer P. H. Internalization of interleukin 1 (IL 1) correlates with IL 1-induced IL 2 receptor expression and IL 2 secretion of EL4 thymoma cells. Eur J Immunol. 1989 Feb;19(2):329–334. doi: 10.1002/eji.1830190217. [DOI] [PubMed] [Google Scholar]
  55. Wijelath E. S., Kardasz A. M., Drummond R., Watson J. Interleukin-one induced inositol phospholipid breakdown in murine macrophages: possible mechanism of receptor activation. Biochem Biophys Res Commun. 1988 Apr 15;152(1):392–397. doi: 10.1016/s0006-291x(88)80726-5. [DOI] [PubMed] [Google Scholar]
  56. Wingfield P., Payton M., Graber P., Rose K., Dayer J. M., Shaw A. R., Schmeissner U. Purification and characterization of human interleukin-1 alpha produced in Escherichia coli. Eur J Biochem. 1987 Jun 15;165(3):537–541. doi: 10.1111/j.1432-1033.1987.tb11472.x. [DOI] [PubMed] [Google Scholar]
  57. Yankner B. A., Shooter E. M. Nerve growth factor in the nucleus: interaction with receptors on the nuclear membrane. Proc Natl Acad Sci U S A. 1979 Mar;76(3):1269–1273. doi: 10.1073/pnas.76.3.1269. [DOI] [PMC free article] [PubMed] [Google Scholar]
  58. Yoneda Y., Imamoto-Sonobe N., Matsuoka Y., Iwamoto R., Kiho Y., Uchida T. Antibodies to Asp-Asp-Glu-Asp can inhibit transport of nuclear proteins into the nucleus. Science. 1988 Oct 14;242(4876):275–278. doi: 10.1126/science.3051382. [DOI] [PubMed] [Google Scholar]
  59. Zeitlin P. L., Hubbard A. L. Cell surface distribution and intracellular fate of asialoglycoproteins: a morphological and biochemical study of isolated rat hepatocytes and monolayer cultures. J Cell Biol. 1982 Mar;92(3):634–647. doi: 10.1083/jcb.92.3.634. [DOI] [PMC free article] [PubMed] [Google Scholar]
  60. Zhang Y. H., Lin J. X., Yip Y. K., Vilcek J. Enhancement of cAMP levels and of protein kinase activity by tumor necrosis factor and interleukin 1 in human fibroblasts: role in the induction of interleukin 6. Proc Natl Acad Sci U S A. 1988 Sep;85(18):6802–6805. doi: 10.1073/pnas.85.18.6802. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES