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. 1999 Apr 15;18(8):2119–2126. doi: 10.1093/emboj/18.8.2119

A casein kinase I motif present in the cytoplasmic domain of members of the tumour necrosis factor ligand family is implicated in 'reverse signalling'.

A D Watts 1, N H Hunt 1, Y Wanigasekara 1, G Bloomfield 1, D Wallach 1, B D Roufogalis 1, G Chaudhri 1
PMCID: PMC1171296  PMID: 10205166

Abstract

We have identified a putative signalling feature of the cytoplasmic domains of the tumour necrosis factor (TNF) family members based on available amino acid sequence data. A casein kinase I (CKI) consensus sequence is conserved in the cytoplasmic domain of six of 15 members of the type II integral membrane TNF ligand family. We examined the phosphorylation state of transmembrane tumour necrosis factor-alpha (mTNF) with [32P]orthophosphate labelling and in vitro kinase assays, in lipopolysaccharide-stimulated RAW264.7 cells. A dimeric form of the type I soluble TNF receptor (sTNFR) was found to dephosphorylate mTNF. This effect could be prevented by treatment with phosphatase inhibitors. Recombinant CKI was able to phosphorylate mTNF that had been dephosphorylated by sTNFR ligation in vivo, and this was less effective if phosphatase inhibitors had been used to prevent mTNF dephosphorylation. A mutated form of mTNF, lacking the CKI recognition site, cannot be phosphorylated by the enzyme. Binding of sTNFR to mTNF induced an increase in intracellular calcium levels in RAW264.7 cells, implying the presence of an associated signalling pathway. We predict that this CKI motif is phosphorylated in other TNF ligand members, and that it represents a new insight into the mechanism of 'reverse signalling' in this cytokine family.

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Selected References

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  1. Aderka D., Englemann H., Hornik V., Skornick Y., Levo Y., Wallach D., Kushtai G. Increased serum levels of soluble receptors for tumor necrosis factor in cancer patients. Cancer Res. 1991 Oct 15;51(20):5602–5607. [PubMed] [Google Scholar]
  2. Bazzoni F., Beutler B. The tumor necrosis factor ligand and receptor families. N Engl J Med. 1996 Jun 27;334(26):1717–1725. doi: 10.1056/NEJM199606273342607. [DOI] [PubMed] [Google Scholar]
  3. Beyaert R., Vanhaesebroeck B., Declercq W., Van Lint J., Vandenabele P., Agostinis P., Vandenheede J. R., Fiers W. Casein kinase-1 phosphorylates the p75 tumor necrosis factor receptor and negatively regulates tumor necrosis factor signaling for apoptosis. J Biol Chem. 1995 Oct 6;270(40):23293–23299. doi: 10.1074/jbc.270.40.23293. [DOI] [PubMed] [Google Scholar]
  4. Cayabyab M., Phillips J. H., Lanier L. L. CD40 preferentially costimulates activation of CD4+ T lymphocytes. J Immunol. 1994 Feb 15;152(4):1523–1531. [PubMed] [Google Scholar]
  5. Chaudhri G. Differential regulation of biosynthesis of cell surface and secreted TNF-alpha in LPS-stimulated murine macrophages. J Leukoc Biol. 1997 Aug;62(2):249–257. doi: 10.1002/jlb.62.2.249. [DOI] [PubMed] [Google Scholar]
  6. Chijiwa T., Hagiwara M., Hidaka H. A newly synthesized selective casein kinase I inhibitor, N-(2-aminoethyl)-5-chloroisoquinoline-8-sulfonamide, and affinity purification of casein kinase I from bovine testis. J Biol Chem. 1989 Mar 25;264(9):4924–4927. [PubMed] [Google Scholar]
  7. Crowe P. D., VanArsdale T. L., Walter B. N., Ware C. F., Hession C., Ehrenfels B., Browning J. L., Din W. S., Goodwin R. G., Smith C. A. A lymphotoxin-beta-specific receptor. Science. 1994 Apr 29;264(5159):707–710. doi: 10.1126/science.8171323. [DOI] [PubMed] [Google Scholar]
  8. Darnay B. G., Singh S., Aggarwal B. B. The p80 TNF receptor-associated kinase (p80TRAK) associates with residues 354-397 of the p80 cytoplasmic domain: similarity to casein kinase. FEBS Lett. 1997 Apr 7;406(1-2):101–105. doi: 10.1016/s0014-5793(97)00251-2. [DOI] [PubMed] [Google Scholar]
  9. Delbridge M. L., Harry J. L., Toder R., O'Neill R. J., Ma K., Chandley A. C., Graves J. A. A human candidate spermatogenesis gene, RBM1, is conserved and amplified on the marsupial Y chromosome. Nat Genet. 1997 Feb;15(2):131–136. doi: 10.1038/ng0297-131. [DOI] [PubMed] [Google Scholar]
  10. Doherty G. M., Lange J. R., Langstein H. N., Alexander H. R., Buresh C. M., Norton J. A. Evidence for IFN-gamma as a mediator of the lethality of endotoxin and tumor necrosis factor-alpha. J Immunol. 1992 Sep 1;149(5):1666–1670. [PubMed] [Google Scholar]
  11. Falini B., Pileri S., Pizzolo G., Dürkop H., Flenghi L., Stirpe F., Martelli M. F., Stein H. CD30 (Ki-1) molecule: a new cytokine receptor of the tumor necrosis factor receptor superfamily as a tool for diagnosis and immunotherapy. Blood. 1995 Jan 1;85(1):1–14. [PubMed] [Google Scholar]
  12. Flotow H., Graves P. R., Wang A. Q., Fiol C. J., Roeske R. W., Roach P. J. Phosphate groups as substrate determinants for casein kinase I action. J Biol Chem. 1990 Aug 25;265(24):14264–14269. [PubMed] [Google Scholar]
  13. Flotow H., Roach P. J. Role of acidic residues as substrate determinants for casein kinase I. J Biol Chem. 1991 Feb 25;266(6):3724–3727. [PubMed] [Google Scholar]
  14. Flotow H., Roach P. J. Synergistic phosphorylation of rabbit muscle glycogen synthase by cyclic AMP-dependent protein kinase and casein kinase I. Implications for hormonal regulation of glycogen synthase. J Biol Chem. 1989 Jun 5;264(16):9126–9128. [PubMed] [Google Scholar]
  15. Gearing A. J., Beckett P., Christodoulou M., Churchill M., Clements J. M., Crimmin M., Davidson A. H., Drummond A. H., Galloway W. A., Gilbert R. Matrix metalloproteinases and processing of pro-TNF-alpha. J Leukoc Biol. 1995 May;57(5):774–777. doi: 10.1002/jlb.57.5.774. [DOI] [PubMed] [Google Scholar]
  16. Gearing A. J., Beckett P., Christodoulou M., Churchill M., Clements J., Davidson A. H., Drummond A. H., Galloway W. A., Gilbert R., Gordon J. L. Processing of tumour necrosis factor-alpha precursor by metalloproteinases. Nature. 1994 Aug 18;370(6490):555–557. doi: 10.1038/370555a0. [DOI] [PubMed] [Google Scholar]
  17. Girardin E., Roux-Lombard P., Grau G. E., Suter P., Gallati H., Dayer J. M. Imbalance between tumour necrosis factor-alpha and soluble TNF receptor concentrations in severe meningococcaemia. The J5 Study Group. Immunology. 1992 May;76(1):20–23. [PMC free article] [PubMed] [Google Scholar]
  18. Godfried M. H., van der Poll T., Jansen J., Romijin J. A., Schattenkerk J. K., Endert E., van Deventer S. J., Sauerwein H. P. Soluble receptors for tumour necrosis factor: a putative marker of disease progression in HIV infection. AIDS. 1993 Jan;7(1):33–36. [PubMed] [Google Scholar]
  19. Graves P. R., Haas D. W., Hagedorn C. H., DePaoli-Roach A. A., Roach P. J. Molecular cloning, expression, and characterization of a 49-kilodalton casein kinase I isoform from rat testis. J Biol Chem. 1993 Mar 25;268(9):6394–6401. [PubMed] [Google Scholar]
  20. Grell M., Douni E., Wajant H., Löhden M., Clauss M., Maxeiner B., Georgopoulos S., Lesslauer W., Kollias G., Pfizenmaier K. The transmembrane form of tumor necrosis factor is the prime activating ligand of the 80 kDa tumor necrosis factor receptor. Cell. 1995 Dec 1;83(5):793–802. doi: 10.1016/0092-8674(95)90192-2. [DOI] [PubMed] [Google Scholar]
  21. Higuchi M., Nagasawa K., Horiuchi T., Oike M., Ito Y., Yasukawa M., Niho Y. Membrane tumor necrosis factor-alpha (TNF-alpha) expressed on HTLV-I-infected T cells mediates a costimulatory signal for B cell activation--characterization of membrane TNF-alpha. Clin Immunol Immunopathol. 1997 Feb;82(2):133–140. doi: 10.1006/clin.1996.4291. [DOI] [PubMed] [Google Scholar]
  22. Janke A., Xu X., Arnason U. The complete mitochondrial genome of the wallaroo (Macropus robustus) and the phylogenetic relationship among Monotremata, Marsupialia, and Eutheria. Proc Natl Acad Sci U S A. 1997 Feb 18;94(4):1276–1281. doi: 10.1073/pnas.94.4.1276. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Jue D. M., Sherry B., Luedke C., Manogue K. R., Cerami A. Processing of newly synthesized cachectin/tumor necrosis factor in endotoxin-stimulated macrophages. Biochemistry. 1990 Sep 11;29(36):8371–8377. doi: 10.1021/bi00488a025. [DOI] [PubMed] [Google Scholar]
  24. Kalinkovich A., Geleziunas R., Kemper O., Belenki D., Wallach D., Wainberg M. A., Bentwich Z. Increased soluble tumor necrosis factor receptor expression and release by human immunodeficiency virus type 1 infection. J Interferon Cytokine Res. 1995 Sep;15(9):749–757. doi: 10.1089/jir.1995.15.749. [DOI] [PubMed] [Google Scholar]
  25. Kennelly P. J., Krebs E. G. Consensus sequences as substrate specificity determinants for protein kinases and protein phosphatases. J Biol Chem. 1991 Aug 25;266(24):15555–15558. [PubMed] [Google Scholar]
  26. Klein J. B., Schepers T. M., Dean W. L., Sonnenfeld G., McLeish K. R. Role of intracellular calcium concentration and protein kinase C activation in IFN-gamma stimulation of U937 cells. J Immunol. 1990 Jun 1;144(11):4305–4311. [PubMed] [Google Scholar]
  27. Kriegler M., Perez C., DeFay K., Albert I., Lu S. D. A novel form of TNF/cachectin is a cell surface cytotoxic transmembrane protein: ramifications for the complex physiology of TNF. Cell. 1988 Apr 8;53(1):45–53. doi: 10.1016/0092-8674(88)90486-2. [DOI] [PubMed] [Google Scholar]
  28. McGeehan G. M., Becherer J. D., Bast R. C., Jr, Boyer C. M., Champion B., Connolly K. M., Conway J. G., Furdon P., Karp S., Kidao S. Regulation of tumour necrosis factor-alpha processing by a metalloproteinase inhibitor. Nature. 1994 Aug 18;370(6490):558–561. doi: 10.1038/370558a0. [DOI] [PubMed] [Google Scholar]
  29. Meggio F., Perich J. W., Marin O., Pinna L. A. The comparative efficiencies of the Ser(P)-, Thr(P)- and Tyr(P)-residues as specificity determinants for casein kinase-1. Biochem Biophys Res Commun. 1992 Feb 14;182(3):1460–1465. doi: 10.1016/0006-291x(92)91898-z. [DOI] [PubMed] [Google Scholar]
  30. Mohler K. M., Sleath P. R., Fitzner J. N., Cerretti D. P., Alderson M., Kerwar S. S., Torrance D. S., Otten-Evans C., Greenstreet T., Weerawarna K. Protection against a lethal dose of endotoxin by an inhibitor of tumour necrosis factor processing. Nature. 1994 Jul 21;370(6486):218–220. doi: 10.1038/370218a0. [DOI] [PubMed] [Google Scholar]
  31. Molyneux M. E., Engelmann H., Taylor T. E., Wirima J. J., Aderka D., Wallach D., Grau G. E. Circulating plasma receptors for tumour necrosis factor in Malawian children with severe falciparum malaria. Cytokine. 1993 Nov;5(6):604–609. doi: 10.1016/s1043-4666(05)80011-0. [DOI] [PubMed] [Google Scholar]
  32. Perez C., Albert I., DeFay K., Zachariades N., Gooding L., Kriegler M. A nonsecretable cell surface mutant of tumor necrosis factor (TNF) kills by cell-to-cell contact. Cell. 1990 Oct 19;63(2):251–258. doi: 10.1016/0092-8674(90)90158-b. [DOI] [PubMed] [Google Scholar]
  33. Pollok K. E., Kim Y. J., Hurtado J., Zhou Z., Kim K. K., Kwon B. S. 4-1BB T-cell antigen binds to mature B cells and macrophages, and costimulates anti-mu-primed splenic B cells. Eur J Immunol. 1994 Feb;24(2):367–374. doi: 10.1002/eji.1830240215. [DOI] [PubMed] [Google Scholar]
  34. Pócsik E., Duda E., Wallach D. Phosphorylation of the 26 kDa TNF precursor in monocytic cells and in transfected HeLa cells. J Inflamm. 1995;45(3):152–160. [PubMed] [Google Scholar]
  35. Ralph P., Nakoinz I. Antibody-dependent killing of erythrocyte and tumor targets by macrophage-related cell lines: enhancement by PPD and LPS. J Immunol. 1977 Sep;119(3):950–954. [PubMed] [Google Scholar]
  36. Raschke W. C., Baird S., Ralph P., Nakoinz I. Functional macrophage cell lines transformed by Abelson leukemia virus. Cell. 1978 Sep;15(1):261–267. doi: 10.1016/0092-8674(78)90101-0. [DOI] [PubMed] [Google Scholar]
  37. Rivers A., Gietzen K. F., Vielhaber E., Virshup D. M. Regulation of casein kinase I epsilon and casein kinase I delta by an in vivo futile phosphorylation cycle. J Biol Chem. 1998 Jun 26;273(26):15980–15984. doi: 10.1074/jbc.273.26.15980. [DOI] [PubMed] [Google Scholar]
  38. Smith C. A., Farrah T., Goodwin R. G. The TNF receptor superfamily of cellular and viral proteins: activation, costimulation, and death. Cell. 1994 Mar 25;76(6):959–962. doi: 10.1016/0092-8674(94)90372-7. [DOI] [PubMed] [Google Scholar]
  39. Stüber E., Neurath M., Calderhead D., Fell H. P., Strober W. Cross-linking of OX40 ligand, a member of the TNF/NGF cytokine family, induces proliferation and differentiation in murine splenic B cells. Immunity. 1995 May;2(5):507–521. doi: 10.1016/1074-7613(95)90031-4. [DOI] [PubMed] [Google Scholar]
  40. Suzuki I., Fink P. J. Maximal proliferation of cytotoxic T lymphocytes requires reverse signaling through Fas ligand. J Exp Med. 1998 Jan 5;187(1):123–128. doi: 10.1084/jem.187.1.123. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Tuazon P. T., Pang D. T., Shafer J. A., Traugh J. A. Phosphorylation of the insulin receptor by casein kinase I. J Cell Biochem. 1985;28(2):159–170. doi: 10.1002/jcb.240280208. [DOI] [PubMed] [Google Scholar]
  42. Tuazon P. T., Traugh J. A. Casein kinase I and II--multipotential serine protein kinases: structure, function, and regulation. Adv Second Messenger Phosphoprotein Res. 1991;23:123–164. [PubMed] [Google Scholar]
  43. Utsumi T., Akimaru K., Kawabata Z., Levitan A., Tokunaga T., Tang P., Ide A., Hung M. C., Klostergaard J. Human pro-tumor necrosis factor: molecular determinants of membrane translocation, sorting, and maturation. Mol Cell Biol. 1995 Nov;15(11):6398–6405. doi: 10.1128/mcb.15.11.6398. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Waser M., Maggiorini M., Lüthy A., Laske A., von Segesser L., Mohacsi P., Opravil M., Turina M., Follath F., Gallino A. Infectious complications in 100 consecutive heart transplant recipients. Eur J Clin Microbiol Infect Dis. 1994 Jan;13(1):12–18. doi: 10.1007/BF02026117. [DOI] [PubMed] [Google Scholar]
  45. Watts A. D., Hunt N. H., Hambly B. D., Chaudhri G. Separation of tumor necrosis factor alpha isoforms by two-dimensional polyacrylamide gel electrophoresis. Electrophoresis. 1997 Jun;18(7):1086–1091. doi: 10.1002/elps.1150180710. [DOI] [PubMed] [Google Scholar]
  46. Wedlock D. N., Aldwell F. E., Buddle B. M. Molecular cloning and characterization of tumor necrosis factor alpha (TNF-alpha) from the Australian common brushtail possum, Trichosurus vulpecula. Immunol Cell Biol. 1996 Apr;74(2):151–158. doi: 10.1038/icb.1996.20. [DOI] [PubMed] [Google Scholar]
  47. Wiley S. R., Goodwin R. G., Smith C. A. Reverse signaling via CD30 ligand. J Immunol. 1996 Oct 15;157(8):3635–3639. [PubMed] [Google Scholar]
  48. Zhang J., Gross S. D., Schroeder M. D., Anderson R. A. Casein kinase I alpha and alpha L: alternative splicing-generated kinases exhibit different catalytic properties. Biochemistry. 1996 Dec 17;35(50):16319–16327. doi: 10.1021/bi9614444. [DOI] [PubMed] [Google Scholar]
  49. de Gunzburg J., Riehl R., Weinberg R. A. Identification of a protein associated with p21ras by chemical crosslinking. Proc Natl Acad Sci U S A. 1989 Jun;86(11):4007–4011. doi: 10.1073/pnas.86.11.4007. [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. van Essen D., Kikutani H., Gray D. CD40 ligand-transduced co-stimulation of T cells in the development of helper function. Nature. 1995 Dec 7;378(6557):620–623. doi: 10.1038/378620a0. [DOI] [PubMed] [Google Scholar]

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