Identification of the cytolinker plectin as a major early in vivo substrate for caspase 8 during CD95- and tumor necrosis factor receptor- mediated apoptosis

Alexander H. Stegh, Harald Herrmann, Stefan Lampel, Dieter Weisenberger, Kerstin Andrä, Martin Seper, Gerhard Wiche, Peter H. Krammer, Marcus E. Peter*

*Corresponding author for this work

Research output: Contribution to journalArticle

128 Citations (Scopus)

Abstract

Caspase 8 plays an essential role in the execution of death receptor- mediated apoptosis. To determine the localization of endogenous caspase 8, we used a panel of subunit-specific anti-caspase 8 monoclonal antibodies in confocal immunofluorescence microscopy. In the human breast carcinoma cell line MCF7, caspase 8 predominantly colocalized with and bound to mitochondria. After induction of apoptosis through CD95 or tumor necrosis factor receptor I, active caspase 8 translocated to plectin, a major cross- linking protein of the three main cytoplasmic filament systems, whereas the caspase 8 prodomain remained bound to mitochondria. Plectin was quantitatively cleaved by caspase 8 at Asp 2395 in the center of the molecule in all cells tested. Cleavage of plectin clearly preceded that of other caspase substrates such as poly(ADP-ribose) polymerase, gelsolin, cytokeratins, or lamin B. In primary fibroblasts from plectin-deficient mice, apoptosis-induced reorganization of the actin cytoskeleton, as seen in wild- type cells, was severely impaired, suggesting that during apoptosis, plectin is required for the reorganization of the microfilament system.

Original languageEnglish (US)
Pages (from-to)5665-5679
Number of pages15
JournalMolecular and Cellular Biology
Volume20
Issue number15
DOIs
StatePublished - Aug 1 2000

Fingerprint

Plectin
Caspase 8
Tumor Necrosis Factor Receptors
Apoptosis
Actin Cytoskeleton
Mitochondria
Lamin Type B
Gelsolin
Death Domain Receptors
Poly(ADP-ribose) Polymerases
Caspases
Keratins
Cytoskeleton
Fluorescence Microscopy
Confocal Microscopy
Fibroblasts
Monoclonal Antibodies
Breast Neoplasms
Cell Line

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology

Cite this

Stegh, Alexander H. ; Herrmann, Harald ; Lampel, Stefan ; Weisenberger, Dieter ; Andrä, Kerstin ; Seper, Martin ; Wiche, Gerhard ; Krammer, Peter H. ; Peter, Marcus E. / Identification of the cytolinker plectin as a major early in vivo substrate for caspase 8 during CD95- and tumor necrosis factor receptor- mediated apoptosis. In: Molecular and Cellular Biology. 2000 ; Vol. 20, No. 15. pp. 5665-5679.
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Identification of the cytolinker plectin as a major early in vivo substrate for caspase 8 during CD95- and tumor necrosis factor receptor- mediated apoptosis. / Stegh, Alexander H.; Herrmann, Harald; Lampel, Stefan; Weisenberger, Dieter; Andrä, Kerstin; Seper, Martin; Wiche, Gerhard; Krammer, Peter H.; Peter, Marcus E.

In: Molecular and Cellular Biology, Vol. 20, No. 15, 01.08.2000, p. 5665-5679.

Research output: Contribution to journalArticle

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AU - Stegh, Alexander H.

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AU - Andrä, Kerstin

AU - Seper, Martin

AU - Wiche, Gerhard

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AU - Peter, Marcus E.

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N2 - Caspase 8 plays an essential role in the execution of death receptor- mediated apoptosis. To determine the localization of endogenous caspase 8, we used a panel of subunit-specific anti-caspase 8 monoclonal antibodies in confocal immunofluorescence microscopy. In the human breast carcinoma cell line MCF7, caspase 8 predominantly colocalized with and bound to mitochondria. After induction of apoptosis through CD95 or tumor necrosis factor receptor I, active caspase 8 translocated to plectin, a major cross- linking protein of the three main cytoplasmic filament systems, whereas the caspase 8 prodomain remained bound to mitochondria. Plectin was quantitatively cleaved by caspase 8 at Asp 2395 in the center of the molecule in all cells tested. Cleavage of plectin clearly preceded that of other caspase substrates such as poly(ADP-ribose) polymerase, gelsolin, cytokeratins, or lamin B. In primary fibroblasts from plectin-deficient mice, apoptosis-induced reorganization of the actin cytoskeleton, as seen in wild- type cells, was severely impaired, suggesting that during apoptosis, plectin is required for the reorganization of the microfilament system.

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