Skip to main content
PMC home page
The EMBO Journal logoLink to The EMBO Journal
. 1998 Oct 15;17(20):5974–5986. doi: 10.1093/emboj/17.20.5974

DEDD, a novel death effector domain-containing protein, targeted to the nucleolus.

A H Stegh 1, O Schickling 1, A Ehret 1, C Scaffidi 1, C Peterhänsel 1, T G Hofmann 1, I Grummt 1, P H Krammer 1, M E Peter 1
PMCID: PMC1170924  PMID: 9774341

Abstract

The CD95 signaling pathway comprises proteins that contain one or two death effector domains (DED), such as FADD/Mort1 or caspase-8. Here we describe a novel 37 kDa protein, DEDD, that contains an N-terminal DED. DEDD is highly conserved between human and mouse (98. 7% identity) and is ubiquitously expressed. Overexpression of DEDD in 293T cells induced weak apoptosis, mainly through its DED by which it interacts with FADD and caspase-8. Endogenous DEDD was found in the cytoplasm and translocated into the nucleus upon stimulation of CD95. Immunocytological studies revealed that overexpressed DEDD directly translocated into the nucleus, where it co-localizes in the nucleolus with UBF, a basal factor required for RNA polymerase I transcription. Consistent with its nuclear localization, DEDD contains two nuclear localization signals and the C-terminal part shares sequence homology with histones. Recombinant DEDD binds to both DNA and reconstituted mononucleosomes and inhibits transcription in a reconstituted in vitro system. The results suggest that DEDD is a final target of a chain of events by which the CD95-induced apoptotic signal is transferred into the nucleolus to shut off cellular biosynthetic activities.

Full Text

The Full Text of this article is available as a PDF (1,006.8 KB).

Selected References

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

  1. Becker P. B. The establishment of active promoters in chromatin. Bioessays. 1994 Aug;16(8):541–547. doi: 10.1002/bies.950160807. [DOI] [PubMed] [Google Scholar]
  2. Boldin M. P., Goncharov T. M., Goltsev Y. V., Wallach D. Involvement of MACH, a novel MORT1/FADD-interacting protease, in Fas/APO-1- and TNF receptor-induced cell death. Cell. 1996 Jun 14;85(6):803–815. doi: 10.1016/s0092-8674(00)81265-9. [DOI] [PubMed] [Google Scholar]
  3. Boldin M. P., Varfolomeev E. E., Pancer Z., Mett I. L., Camonis J. H., Wallach D. A novel protein that interacts with the death domain of Fas/APO1 contains a sequence motif related to the death domain. J Biol Chem. 1995 Apr 7;270(14):7795–7798. doi: 10.1074/jbc.270.14.7795. [DOI] [PubMed] [Google Scholar]
  4. Casiano C. A., Martin S. J., Green D. R., Tan E. M. Selective cleavage of nuclear autoantigens during CD95 (Fas/APO-1)-mediated T cell apoptosis. J Exp Med. 1996 Aug 1;184(2):765–770. doi: 10.1084/jem.184.2.765. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Casiano C. A., Ochs R. L., Tan E. M. Distinct cleavage products of nuclear proteins in apoptosis and necrosis revealed by autoantibody probes. Cell Death Differ. 1998 Feb;5(2):183–190. doi: 10.1038/sj.cdd.4400336. [DOI] [PubMed] [Google Scholar]
  6. Casiano C. A., Tan E. M. Antinuclear autoantibodies: probes for defining proteolytic events associated with apoptosis. Mol Biol Rep. 1996;23(3-4):211–216. doi: 10.1007/BF00351171. [DOI] [PubMed] [Google Scholar]
  7. Chan E. K., Imai H., Hamel J. C., Tan E. M. Human autoantibody to RNA polymerase I transcription factor hUBF. Molecular identity of nucleolus organizer region autoantigen NOR-90 and ribosomal RNA transcription upstream binding factor. J Exp Med. 1991 Nov 1;174(5):1239–1244. doi: 10.1084/jem.174.5.1239. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Chinnaiyan A. M., O'Rourke K., Tewari M., Dixit V. M. FADD, a novel death domain-containing protein, interacts with the death domain of Fas and initiates apoptosis. Cell. 1995 May 19;81(4):505–512. doi: 10.1016/0092-8674(95)90071-3. [DOI] [PubMed] [Google Scholar]
  9. Chinnaiyan A. M., Tepper C. G., Seldin M. F., O'Rourke K., Kischkel F. C., Hellbardt S., Krammer P. H., Peter M. E., Dixit V. M. FADD/MORT1 is a common mediator of CD95 (Fas/APO-1) and tumor necrosis factor receptor-induced apoptosis. J Biol Chem. 1996 Mar 1;271(9):4961–4965. doi: 10.1074/jbc.271.9.4961. [DOI] [PubMed] [Google Scholar]
  10. Croston G. E., Kerrigan L. A., Lira L. M., Marshak D. R., Kadonaga J. T. Sequence-specific antirepression of histone H1-mediated inhibition of basal RNA polymerase II transcription. Science. 1991 Feb 8;251(4994):643–649. doi: 10.1126/science.1899487. [DOI] [PubMed] [Google Scholar]
  11. Eberstadt M., Huang B., Chen Z., Meadows R. P., Ng S. C., Zheng L., Lenardo M. J., Fesik S. W. NMR structure and mutagenesis of the FADD (Mort1) death-effector domain. Nature. 1998 Apr 30;392(6679):941–945. doi: 10.1038/31972. [DOI] [PubMed] [Google Scholar]
  12. Enari M., Sakahira H., Yokoyama H., Okawa K., Iwamatsu A., Nagata S. A caspase-activated DNase that degrades DNA during apoptosis, and its inhibitor ICAD. Nature. 1998 Jan 1;391(6662):43–50. doi: 10.1038/34112. [DOI] [PubMed] [Google Scholar]
  13. Fernandes-Alnemri T., Armstrong R. C., Krebs J., Srinivasula S. M., Wang L., Bullrich F., Fritz L. C., Trapani J. A., Tomaselli K. J., Litwack G. In vitro activation of CPP32 and Mch3 by Mch4, a novel human apoptotic cysteine protease containing two FADD-like domains. Proc Natl Acad Sci U S A. 1996 Jul 23;93(15):7464–7469. doi: 10.1073/pnas.93.15.7464. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Goltsev Y. V., Kovalenko A. V., Arnold E., Varfolomeev E. E., Brodianskii V. M., Wallach D. CASH, a novel caspase homologue with death effector domains. J Biol Chem. 1997 Aug 8;272(32):19641–19644. doi: 10.1074/jbc.272.32.19641. [DOI] [PubMed] [Google Scholar]
  15. Han D. K., Chaudhary P. M., Wright M. E., Friedman C., Trask B. J., Riedel R. T., Baskin D. G., Schwartz S. M., Hood L. MRIT, a novel death-effector domain-containing protein, interacts with caspases and BclXL and initiates cell death. Proc Natl Acad Sci U S A. 1997 Oct 14;94(21):11333–11338. doi: 10.1073/pnas.94.21.11333. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Houge G., Robaye B., Eikhom T. S., Golstein J., Mellgren G., Gjertsen B. T., Lanotte M., Døskeland S. O. Fine mapping of 28S rRNA sites specifically cleaved in cells undergoing apoptosis. Mol Cell Biol. 1995 Apr;15(4):2051–2062. doi: 10.1128/mcb.15.4.2051. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Hsu Y. T., Wolter K. G., Youle R. J. Cytosol-to-membrane redistribution of Bax and Bcl-X(L) during apoptosis. Proc Natl Acad Sci U S A. 1997 Apr 15;94(8):3668–3672. doi: 10.1073/pnas.94.8.3668. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Hu S., Vincenz C., Ni J., Gentz R., Dixit V. M. I-FLICE, a novel inhibitor of tumor necrosis factor receptor-1- and CD-95-induced apoptosis. J Biol Chem. 1997 Jul 11;272(28):17255–17257. doi: 10.1074/jbc.272.28.17255. [DOI] [PubMed] [Google Scholar]
  19. Inohara N., Koseki T., Hu Y., Chen S., Núez G. CLARP, a death effector domain-containing protein interacts with caspase-8 and regulates apoptosis. Proc Natl Acad Sci U S A. 1997 Sep 30;94(20):10717–10722. doi: 10.1073/pnas.94.20.10717. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Irmler M., Thome M., Hahne M., Schneider P., Hofmann K., Steiner V., Bodmer J. L., Schröter M., Burns K., Mattmann C. Inhibition of death receptor signals by cellular FLIP. Nature. 1997 Jul 10;388(6638):190–195. doi: 10.1038/40657. [DOI] [PubMed] [Google Scholar]
  21. Kischkel F. C., Hellbardt S., Behrmann I., Germer M., Pawlita M., Krammer P. H., Peter M. E. Cytotoxicity-dependent APO-1 (Fas/CD95)-associated proteins form a death-inducing signaling complex (DISC) with the receptor. EMBO J. 1995 Nov 15;14(22):5579–5588. doi: 10.1002/j.1460-2075.1995.tb00245.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Kuhn A., Grummt I. Dual role of the nucleolar transcription factor UBF: trans-activator and antirepressor. Proc Natl Acad Sci U S A. 1992 Aug 15;89(16):7340–7344. doi: 10.1073/pnas.89.16.7340. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Liu X., Zou H., Slaughter C., Wang X. DFF, a heterodimeric protein that functions downstream of caspase-3 to trigger DNA fragmentation during apoptosis. Cell. 1997 Apr 18;89(2):175–184. doi: 10.1016/s0092-8674(00)80197-x. [DOI] [PubMed] [Google Scholar]
  24. Luger K., Richmond T. J. DNA binding within the nucleosome core. Curr Opin Struct Biol. 1998 Feb;8(1):33–40. doi: 10.1016/s0959-440x(98)80007-9. [DOI] [PubMed] [Google Scholar]
  25. Längst G., Becker P. B., Grummt I. TTF-I determines the chromatin architecture of the active rDNA promoter. EMBO J. 1998 Jun 1;17(11):3135–3145. doi: 10.1093/emboj/17.11.3135. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Längst G., Blank T. A., Becker P. B., Grummt I. RNA polymerase I transcription on nucleosomal templates: the transcription termination factor TTF-I induces chromatin remodeling and relieves transcriptional repression. EMBO J. 1997 Feb 17;16(4):760–768. doi: 10.1093/emboj/16.4.760. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Medema J. P., Scaffidi C., Kischkel F. C., Shevchenko A., Mann M., Krammer P. H., Peter M. E. FLICE is activated by association with the CD95 death-inducing signaling complex (DISC). EMBO J. 1997 May 15;16(10):2794–2804. doi: 10.1093/emboj/16.10.2794. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Medema J. P., Scaffidi C., Krammer P. H., Peter M. E. Bcl-xL acts downstream of caspase-8 activation by the CD95 death-inducing signaling complex. J Biol Chem. 1998 Feb 6;273(6):3388–3393. doi: 10.1074/jbc.273.6.3388. [DOI] [PubMed] [Google Scholar]
  29. Muzio M., Chinnaiyan A. M., Kischkel F. C., O'Rourke K., Shevchenko A., Ni J., Scaffidi C., Bretz J. D., Zhang M., Gentz R. FLICE, a novel FADD-homologous ICE/CED-3-like protease, is recruited to the CD95 (Fas/APO-1) death--inducing signaling complex. Cell. 1996 Jun 14;85(6):817–827. doi: 10.1016/s0092-8674(00)81266-0. [DOI] [PubMed] [Google Scholar]
  30. Peter M. E., Hellbardt S., Schwartz-Albiez R., Westendorp M. O., Walczak H., Moldenhauer G., Grell M., Krammer P. H. Cell surface sialylation plays a role in modulating sensitivity towards APO-1-mediated apoptotic cell death. Cell Death Differ. 1995 Jul;2(3):163–171. [PubMed] [Google Scholar]
  31. Peter M. E., Medema J. P., Krammer P. H. Does the Caenorhabditis elegans protein CED-4 contain a region of homology to the mammalian death effector domain? Cell Death Differ. 1997 Oct;4(7):523–525. doi: 10.1038/sj.cdd.4400285. [DOI] [PubMed] [Google Scholar]
  32. Rasper D. M., Vaillancourt J. P., Hadano S., Houtzager V. M., Seiden I., Keen S. L., Tawa P., Xanthoudakis S., Nasir J., Martindale D. Cell death attenuation by 'Usurpin', a mammalian DED-caspase homologue that precludes caspase-8 recruitment and activation by the CD-95 (Fas, APO-1) receptor complex. Cell Death Differ. 1998 Apr;5(4):271–288. doi: 10.1038/sj.cdd.4400370. [DOI] [PubMed] [Google Scholar]
  33. Renz A., Fackelmayer F. O. Purification and molecular cloning of the scaffold attachment factor B (SAF-B), a novel human nuclear protein that specifically binds to S/MAR-DNA. Nucleic Acids Res. 1996 Mar 1;24(5):843–849. doi: 10.1093/nar/24.5.843. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Rhéaume E., Cohen L. Y., Uhlmann F., Lazure C., Alam A., Hurwitz J., Sékaly R. P., Denis F. The large subunit of replication factor C is a substrate for caspase-3 in vitro and is cleaved by a caspase-3-like protease during Fas-mediated apoptosis. EMBO J. 1997 Nov 3;16(21):6346–6354. doi: 10.1093/emboj/16.21.6346. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Sakahira H., Enari M., Nagata S. Cleavage of CAD inhibitor in CAD activation and DNA degradation during apoptosis. Nature. 1998 Jan 1;391(6662):96–99. doi: 10.1038/34214. [DOI] [PubMed] [Google Scholar]
  36. Scaffidi C., Fulda S., Srinivasan A., Friesen C., Li F., Tomaselli K. J., Debatin K. M., Krammer P. H., Peter M. E. Two CD95 (APO-1/Fas) signaling pathways. EMBO J. 1998 Mar 16;17(6):1675–1687. doi: 10.1093/emboj/17.6.1675. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Scaffidi C., Medema J. P., Krammer P. H., Peter M. E. FLICE is predominantly expressed as two functionally active isoforms, caspase-8/a and caspase-8/b. J Biol Chem. 1997 Oct 24;272(43):26953–26958. doi: 10.1074/jbc.272.43.26953. [DOI] [PubMed] [Google Scholar]
  38. Senger M., Glatting K. H., Ritter O., Suhai S. X-HUSAR, an X-based graphical interface for the analysis of genomic sequences. Comput Methods Programs Biomed. 1995 Feb;46(2):131–141. doi: 10.1016/0169-2607(94)01610-r. [DOI] [PubMed] [Google Scholar]
  39. Shaw P. J., Jordan E. G. The nucleolus. Annu Rev Cell Dev Biol. 1995;11:93–121. doi: 10.1146/annurev.cb.11.110195.000521. [DOI] [PubMed] [Google Scholar]
  40. Shields J. M., Yang V. W. Two potent nuclear localization signals in the gut-enriched Krüppel-like factor define a subfamily of closely related Krüppel proteins. J Biol Chem. 1997 Jul 18;272(29):18504–18507. doi: 10.1074/jbc.272.29.18504. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Shu H. B., Halpin D. R., Goeddel D. V. Casper is a FADD- and caspase-related inducer of apoptosis. Immunity. 1997 Jun;6(6):751–763. doi: 10.1016/s1074-7613(00)80450-1. [DOI] [PubMed] [Google Scholar]
  42. Smith C. M., Steitz J. A. Sno storm in the nucleolus: new roles for myriad small RNPs. Cell. 1997 May 30;89(5):669–672. doi: 10.1016/s0092-8674(00)80247-0. [DOI] [PubMed] [Google Scholar]
  43. Song Q., Lu H., Zhang N., Luckow B., Shah G., Poirier G., Lavin M. Specific cleavage of the large subunit of replication factor C in apoptosis is mediated by CPP32-like protease. Biochem Biophys Res Commun. 1997 Apr 17;233(2):343–348. doi: 10.1006/bbrc.1997.6456. [DOI] [PubMed] [Google Scholar]
  44. Srinivasula S. M., Ahmad M., Ottilie S., Bullrich F., Banks S., Wang Y., Fernandes-Alnemri T., Croce C. M., Litwack G., Tomaselli K. J. FLAME-1, a novel FADD-like anti-apoptotic molecule that regulates Fas/TNFR1-induced apoptosis. J Biol Chem. 1997 Jul 25;272(30):18542–18545. doi: 10.1074/jbc.272.30.18542. [DOI] [PubMed] [Google Scholar]
  45. Stein A. Reconstitution of chromatin from purified components. Methods Enzymol. 1989;170:585–603. doi: 10.1016/0076-6879(89)70066-5. [DOI] [PubMed] [Google Scholar]
  46. Südbeck P., Scherer G. Two independent nuclear localization signals are present in the DNA-binding high-mobility group domains of SRY and SOX9. J Biol Chem. 1997 Oct 31;272(44):27848–27852. doi: 10.1074/jbc.272.44.27848. [DOI] [PubMed] [Google Scholar]
  47. Ubeda M., Habener J. F. The large subunit of the DNA replication complex C (DSEB/RF-C140) cleaved and inactivated by caspase-3 (CPP32/YAMA) during Fas-induced apoptosis. J Biol Chem. 1997 Aug 1;272(31):19562–19568. doi: 10.1074/jbc.272.31.19562. [DOI] [PubMed] [Google Scholar]
  48. Vincenz C., Dixit V. M. Fas-associated death domain protein interleukin-1beta-converting enzyme 2 (FLICE2), an ICE/Ced-3 homologue, is proximally involved in CD95- and p55-mediated death signaling. J Biol Chem. 1997 Mar 7;272(10):6578–6583. doi: 10.1074/jbc.272.10.6578. [DOI] [PubMed] [Google Scholar]
  49. Ylikomi T., Bocquel M. T., Berry M., Gronemeyer H., Chambon P. Cooperation of proto-signals for nuclear accumulation of estrogen and progesterone receptors. EMBO J. 1992 Oct;11(10):3681–3694. doi: 10.1002/j.1460-2075.1992.tb05453.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. Zhou B. B., Li H., Yuan J., Kirschner M. W. Caspase-dependent activation of cyclin-dependent kinases during Fas-induced apoptosis in Jurkat cells. Proc Natl Acad Sci U S A. 1998 Jun 9;95(12):6785–6790. doi: 10.1073/pnas.95.12.6785. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from The EMBO Journal are provided here courtesy of Nature Publishing Group

RESOURCES