In mouse embryonic fibroblasts, neither caspase-8 nor cellular FLICE-inhibitory protein (FLIP) is necessary for TNF to activate NF-kappaB, but caspase-8 is required for TNF to cause cell death, and induction of FLIP by NF-kappaB is required to preven

Donia Moujalled, Wendy Cook, Josep Lluis, Nufail Khan, Afsar Ahmed, Bernard Callus, David Vaux

Research output: Contribution to journal › Article › Research › peer-review

Abstract

Binding of TNF to TNF receptor-1 can give a pro-survival signal through activation of p65/RelA NF-kappaB, but also signals cell death. To determine the roles of FLICE-inhibitory protein (FLIP) and caspase-8 in TNF-induced activation of NF-kappaB and apoptosis, we used mouse embryonic fibroblasts derived from FLIP and caspase-8 gene-deleted mice, and treated them with TNF and a smac-mimetic compound that causes degradation of cellular inhibitor of apoptosis proteins (cIAPs). In cells treated with smac mimetic, TNF and Fas Ligand caused wild-type and FLIP(-/-) MEFs to die, whereas caspase-8(-/-) MEFs survived, indicating that caspase-8 is necessary for death of MEFs triggered by these ligands when IAPs are degraded. By contrast, neither caspase-8 nor FLIP was required for TNF to activate p65/RelA NF-kappaB, because IkappaB was degraded, p65 translocated to the nucleus, and an NF-kappaB reporter gene activated normally in caspase-8(-/-) or FLIP(-/-) MEFs. Reconstitution of FLIP(-/-) MEFs with the FLIP isoforms FLIP-L, FLIP-R, or FLIP-p43 protected these cells from dying when treated with TNF or FasL, whether or not cIAPs were depleted. These results show that in MEFs, caspase-8 is necessary for TNF- and FasL-induced death, and FLIP is needed to prevent it, but neither caspase-8 nor FLIP is required for TNF to activate NF-kappaB.
Original languageEnglish
Pages (from-to)808 - 815
Number of pages8
JournalCell Death and Differentiation
Volume19
Issue number5
DOIs
Publication statusPublished - 2012

Cite this

Moujalled, Donia ; Cook, Wendy ; Lluis, Josep ; Khan, Nufail ; Ahmed, Afsar ; Callus, Bernard ; Vaux, David. / In mouse embryonic fibroblasts, neither caspase-8 nor cellular FLICE-inhibitory protein (FLIP) is necessary for TNF to activate NF-kappaB, but caspase-8 is required for TNF to cause cell death, and induction of FLIP by NF-kappaB is required to preven. In: Cell Death and Differentiation. 2012 ; Vol. 19, No. 5. pp. 808 - 815.
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title = "In mouse embryonic fibroblasts, neither caspase-8 nor cellular FLICE-inhibitory protein (FLIP) is necessary for TNF to activate NF-kappaB, but caspase-8 is required for TNF to cause cell death, and induction of FLIP by NF-kappaB is required to preven",
abstract = "Binding of TNF to TNF receptor-1 can give a pro-survival signal through activation of p65/RelA NF-kappaB, but also signals cell death. To determine the roles of FLICE-inhibitory protein (FLIP) and caspase-8 in TNF-induced activation of NF-kappaB and apoptosis, we used mouse embryonic fibroblasts derived from FLIP and caspase-8 gene-deleted mice, and treated them with TNF and a smac-mimetic compound that causes degradation of cellular inhibitor of apoptosis proteins (cIAPs). In cells treated with smac mimetic, TNF and Fas Ligand caused wild-type and FLIP(-/-) MEFs to die, whereas caspase-8(-/-) MEFs survived, indicating that caspase-8 is necessary for death of MEFs triggered by these ligands when IAPs are degraded. By contrast, neither caspase-8 nor FLIP was required for TNF to activate p65/RelA NF-kappaB, because IkappaB was degraded, p65 translocated to the nucleus, and an NF-kappaB reporter gene activated normally in caspase-8(-/-) or FLIP(-/-) MEFs. Reconstitution of FLIP(-/-) MEFs with the FLIP isoforms FLIP-L, FLIP-R, or FLIP-p43 protected these cells from dying when treated with TNF or FasL, whether or not cIAPs were depleted. These results show that in MEFs, caspase-8 is necessary for TNF- and FasL-induced death, and FLIP is needed to prevent it, but neither caspase-8 nor FLIP is required for TNF to activate NF-kappaB.",
author = "Donia Moujalled and Wendy Cook and Josep Lluis and Nufail Khan and Afsar Ahmed and Bernard Callus and David Vaux",
year = "2012",
doi = "10.1038/cdd.2011.151",
language = "English",
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pages = "808 -- 815",
journal = "Cell Death and Differentiation",
issn = "1350-9047",
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Moujalled, D, Cook, W, Lluis, J, Khan, N, Ahmed, A, Callus, B & Vaux, D 2012, 'In mouse embryonic fibroblasts, neither caspase-8 nor cellular FLICE-inhibitory protein (FLIP) is necessary for TNF to activate NF-kappaB, but caspase-8 is required for TNF to cause cell death, and induction of FLIP by NF-kappaB is required to preven' Cell Death and Differentiation, vol. 19, no. 5, pp. 808 - 815. https://doi.org/10.1038/cdd.2011.151

In mouse embryonic fibroblasts, neither caspase-8 nor cellular FLICE-inhibitory protein (FLIP) is necessary for TNF to activate NF-kappaB, but caspase-8 is required for TNF to cause cell death, and induction of FLIP by NF-kappaB is required to preven. / Moujalled, Donia; Cook, Wendy; Lluis, Josep; Khan, Nufail; Ahmed, Afsar; Callus, Bernard; Vaux, David.

In: Cell Death and Differentiation, Vol. 19, No. 5, 2012, p. 808 - 815.

Research output: Contribution to journal › Article › Research › peer-review

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AU - Moujalled, Donia

AU - Cook, Wendy

AU - Lluis, Josep

AU - Khan, Nufail

AU - Ahmed, Afsar

AU - Callus, Bernard

AU - Vaux, David

PY - 2012

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N2 - Binding of TNF to TNF receptor-1 can give a pro-survival signal through activation of p65/RelA NF-kappaB, but also signals cell death. To determine the roles of FLICE-inhibitory protein (FLIP) and caspase-8 in TNF-induced activation of NF-kappaB and apoptosis, we used mouse embryonic fibroblasts derived from FLIP and caspase-8 gene-deleted mice, and treated them with TNF and a smac-mimetic compound that causes degradation of cellular inhibitor of apoptosis proteins (cIAPs). In cells treated with smac mimetic, TNF and Fas Ligand caused wild-type and FLIP(-/-) MEFs to die, whereas caspase-8(-/-) MEFs survived, indicating that caspase-8 is necessary for death of MEFs triggered by these ligands when IAPs are degraded. By contrast, neither caspase-8 nor FLIP was required for TNF to activate p65/RelA NF-kappaB, because IkappaB was degraded, p65 translocated to the nucleus, and an NF-kappaB reporter gene activated normally in caspase-8(-/-) or FLIP(-/-) MEFs. Reconstitution of FLIP(-/-) MEFs with the FLIP isoforms FLIP-L, FLIP-R, or FLIP-p43 protected these cells from dying when treated with TNF or FasL, whether or not cIAPs were depleted. These results show that in MEFs, caspase-8 is necessary for TNF- and FasL-induced death, and FLIP is needed to prevent it, but neither caspase-8 nor FLIP is required for TNF to activate NF-kappaB.

AB - Binding of TNF to TNF receptor-1 can give a pro-survival signal through activation of p65/RelA NF-kappaB, but also signals cell death. To determine the roles of FLICE-inhibitory protein (FLIP) and caspase-8 in TNF-induced activation of NF-kappaB and apoptosis, we used mouse embryonic fibroblasts derived from FLIP and caspase-8 gene-deleted mice, and treated them with TNF and a smac-mimetic compound that causes degradation of cellular inhibitor of apoptosis proteins (cIAPs). In cells treated with smac mimetic, TNF and Fas Ligand caused wild-type and FLIP(-/-) MEFs to die, whereas caspase-8(-/-) MEFs survived, indicating that caspase-8 is necessary for death of MEFs triggered by these ligands when IAPs are degraded. By contrast, neither caspase-8 nor FLIP was required for TNF to activate p65/RelA NF-kappaB, because IkappaB was degraded, p65 translocated to the nucleus, and an NF-kappaB reporter gene activated normally in caspase-8(-/-) or FLIP(-/-) MEFs. Reconstitution of FLIP(-/-) MEFs with the FLIP isoforms FLIP-L, FLIP-R, or FLIP-p43 protected these cells from dying when treated with TNF or FasL, whether or not cIAPs were depleted. These results show that in MEFs, caspase-8 is necessary for TNF- and FasL-induced death, and FLIP is needed to prevent it, but neither caspase-8 nor FLIP is required for TNF to activate NF-kappaB.

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U2 - 10.1038/cdd.2011.151

DO - 10.1038/cdd.2011.151

M3 - Article

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JO - Cell Death and Differentiation

JF - Cell Death and Differentiation

SN - 1350-9047

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ER -

Moujalled D, Cook W, Lluis J, Khan N, Ahmed A, Callus B et al. In mouse embryonic fibroblasts, neither caspase-8 nor cellular FLICE-inhibitory protein (FLIP) is necessary for TNF to activate NF-kappaB, but caspase-8 is required for TNF to cause cell death, and induction of FLIP by NF-kappaB is required to preven. Cell Death and Differentiation. 2012;19(5):808 - 815. https://doi.org/10.1038/cdd.2011.151

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