DEDD regulates degradation of intermediate filaments during apoptosis

Justine C. Lee, Olaf Schickling, Alexander H. Stegh, Robert G. Oshima, David Dinsdale, Gerald M. Cohen, Marcus E. Peter*

*Corresponding author for this work

Research output: Contribution to journalArticle

66 Citations (Scopus)

Abstract

Apoptosis depends critically on regulated cytoskeletal reorganization events in a cell. We demonstrate that death effector domain containing DNA binding protein (DEDD), a highly conserved and ubiquitous death effector domain containing protein, exists predominantly as mono- or diubiquitinated, and that diubiquitinated DEDD interacts with both the K8/18 intermediate filament network and pro-caspase-3. Early in apoptosis, both cytosolic DEDD and its close homologue DEDD2 formed filaments that colocalized with and depended on K8/18 and active caspase-3. Subsequently, these filamentous structures collapsed into intracellular inclusions that migrated into cytoplasmic blebs and contained DEDD, DEDD2, active caspase-3, and caspase-3-cleaved K18 late in apoptosis. Biochemical studies further confirmed that DEDD coimmunoprecipitated with both K18 and pro-caspase-3, and kinetic analyses placed apoptotic DEDD staining prior to caspase-3 activation and K18 cleavage. In addition, both caspase-3 activation and K18 cleavage was inhibited by expression of DEDDΔNLS1-3, a cytosolic form of DEDD that cannot be ubiquitinated. Finally, siRNA mediated DEDD knockdown cells exhibited inhibition of staurosporine-induced DNA degradation. Our data suggest that DEDD represents a novel scaffold protein that directs the effector caspase-3 to certain substrates facilitating their ordered degradation during apoptosis.

Original languageEnglish (US)
Pages (from-to)1051-1066
Number of pages16
JournalJournal of Cell Biology
Volume158
Issue number6
DOIs
StatePublished - Sep 16 2002

Fingerprint

Intermediate Filaments
Caspase 3
Apoptosis
Death Domain Receptor Signaling Adaptor Proteins
Effector Caspases
Staurosporine
DNA-Binding Proteins
Blister
Small Interfering RNA
Staining and Labeling
K-18 conjugate
DNA

Keywords

  • Apoptosis
  • Caspases
  • DEDD
  • Intermediate filaments
  • Mono-ubiquitination

ASJC Scopus subject areas

  • Cell Biology

Cite this

Lee, Justine C. ; Schickling, Olaf ; Stegh, Alexander H. ; Oshima, Robert G. ; Dinsdale, David ; Cohen, Gerald M. ; Peter, Marcus E. / DEDD regulates degradation of intermediate filaments during apoptosis. In: Journal of Cell Biology. 2002 ; Vol. 158, No. 6. pp. 1051-1066.
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Lee, JC, Schickling, O, Stegh, AH, Oshima, RG, Dinsdale, D, Cohen, GM & Peter, ME 2002, 'DEDD regulates degradation of intermediate filaments during apoptosis', Journal of Cell Biology, vol. 158, no. 6, pp. 1051-1066. https://doi.org/10.1083/jcb.200112124

DEDD regulates degradation of intermediate filaments during apoptosis. / Lee, Justine C.; Schickling, Olaf; Stegh, Alexander H.; Oshima, Robert G.; Dinsdale, David; Cohen, Gerald M.; Peter, Marcus E.

In: Journal of Cell Biology, Vol. 158, No. 6, 16.09.2002, p. 1051-1066.

Research output: Contribution to journalArticle

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T1 - DEDD regulates degradation of intermediate filaments during apoptosis

AU - Lee, Justine C.

AU - Schickling, Olaf

AU - Stegh, Alexander H.

AU - Oshima, Robert G.

AU - Dinsdale, David

AU - Cohen, Gerald M.

AU - Peter, Marcus E.

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AB - Apoptosis depends critically on regulated cytoskeletal reorganization events in a cell. We demonstrate that death effector domain containing DNA binding protein (DEDD), a highly conserved and ubiquitous death effector domain containing protein, exists predominantly as mono- or diubiquitinated, and that diubiquitinated DEDD interacts with both the K8/18 intermediate filament network and pro-caspase-3. Early in apoptosis, both cytosolic DEDD and its close homologue DEDD2 formed filaments that colocalized with and depended on K8/18 and active caspase-3. Subsequently, these filamentous structures collapsed into intracellular inclusions that migrated into cytoplasmic blebs and contained DEDD, DEDD2, active caspase-3, and caspase-3-cleaved K18 late in apoptosis. Biochemical studies further confirmed that DEDD coimmunoprecipitated with both K18 and pro-caspase-3, and kinetic analyses placed apoptotic DEDD staining prior to caspase-3 activation and K18 cleavage. In addition, both caspase-3 activation and K18 cleavage was inhibited by expression of DEDDΔNLS1-3, a cytosolic form of DEDD that cannot be ubiquitinated. Finally, siRNA mediated DEDD knockdown cells exhibited inhibition of staurosporine-induced DNA degradation. Our data suggest that DEDD represents a novel scaffold protein that directs the effector caspase-3 to certain substrates facilitating their ordered degradation during apoptosis.

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Lee JC, Schickling O, Stegh AH, Oshima RG, Dinsdale D, Cohen GM et al. DEDD regulates degradation of intermediate filaments during apoptosis. Journal of Cell Biology. 2002 Sep 16;158(6):1051-1066. https://doi.org/10.1083/jcb.200112124