ERK-1 MAP kinase prevents TNF-induced apoptosis through bad phosphorylation and inhibition of bax translocation in HeLa cells

Bruna Pucci, Manuela Indelicato, Valentina Paradisi, Valentina Reali, Laura Pellegrini, Michele Aventaggiato, Natalie O. Karpinich, Massimo Fini, Matteo A. Russo, John L. Farber, Marco Tafani

Research output: Contribution to journalArticle

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Abstract

Extracellular signal-regulated kinase (ERK) 1/2 signaling is involved in tumor cell survival through the regulation of Bcl-2 family members. To explore this further and to demonstrate the central role of the mitochondria in the ERK1/2 pathway we used the HeLa cellular model where apoptosis was induced by tumor necrosis factor (TNF) and cycloheximide (CHX). We show that HeLa cells overexpressing ERK-1 displayed resistance to TNF and CHX. HeLa cells overexpressing a kinase-deficient form of ERK-1 (K71R) were more sensitive to TNF and CHX. In the ERK-1 cells, Bad was phosphorylated during TNF + CHX treatment. In the HeLa wt cells and in the K71R clones TNF and CHX decreased Bad phosphorylation. ERK-1 cells treated with TNF and CHX did not release cytochrome c from the mitochondria. By contrast, HeLa wt and K71R clones released cytochrome c. Bax did not translocate to the mitochondria in ERK-1 cells treated with TNF + CHX. Conversely, HeLa wt and K71R clones accumulated Bax in the mitochondria. In the HeLa wt cells and in both ERK-1 transfectants Bid was cleaved and accumulated in the mitochondria. The caspase-8 inhibitor IETD-FMK and the mitochondrial membrane permeabilization inhibitor bongkrekic acid (BK), partially prevented cell death by TNF + CHX. Anisomycin, a c-Jun N-terminal kinases activator, increased TNF-killing. The ERK-1 cells were resistant to TNF and anisomycin, whereas K71R clones resulted more sensitive. Our study demonstrates that in HeLa cells the ERK-1 kinase prevents TNF + CHX apoptosis by regulating the intrinsic mitochondrial pathway through different mechanisms. Inhibition of the intrinsic pathway is sufficient to almost completely prevent cell death.

Original languageEnglish
Pages (from-to)1166-1174
Number of pages9
JournalJournal of Cellular Biochemistry
Volume108
Issue number5
DOIs
Publication statusPublished - Dec 1 2009

Fingerprint

Phosphorylation
Mitogen-Activated Protein Kinase 3
Mitogen-Activated Protein Kinase Kinases
Extracellular Signal-Regulated MAP Kinases
HeLa Cells
Cycloheximide
Phosphotransferases
Tumor Necrosis Factor-alpha
Apoptosis
Mitochondria
Clone Cells
Anisomycin
Cell death
Cytochromes c
Bongkrekic Acid
Inhibition (Psychology)
Cell Death
Caspase Inhibitors
Caspase 8
MAP Kinase Signaling System

Keywords

  • Anisomycin
  • Bad phosphorylation
  • Bax
  • Erk-1
  • TNF

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Molecular Biology

Cite this

Pucci, B., Indelicato, M., Paradisi, V., Reali, V., Pellegrini, L., Aventaggiato, M., ... Tafani, M. (2009). ERK-1 MAP kinase prevents TNF-induced apoptosis through bad phosphorylation and inhibition of bax translocation in HeLa cells. Journal of Cellular Biochemistry, 108(5), 1166-1174. https://doi.org/10.1002/jcb.22345

ERK-1 MAP kinase prevents TNF-induced apoptosis through bad phosphorylation and inhibition of bax translocation in HeLa cells. / Pucci, Bruna; Indelicato, Manuela; Paradisi, Valentina; Reali, Valentina; Pellegrini, Laura; Aventaggiato, Michele; Karpinich, Natalie O.; Fini, Massimo; Russo, Matteo A.; Farber, John L.; Tafani, Marco.

In: Journal of Cellular Biochemistry, Vol. 108, No. 5, 01.12.2009, p. 1166-1174.

Research output: Contribution to journalArticle

Pucci, B, Indelicato, M, Paradisi, V, Reali, V, Pellegrini, L, Aventaggiato, M, Karpinich, NO, Fini, M, Russo, MA, Farber, JL & Tafani, M 2009, 'ERK-1 MAP kinase prevents TNF-induced apoptosis through bad phosphorylation and inhibition of bax translocation in HeLa cells', Journal of Cellular Biochemistry, vol. 108, no. 5, pp. 1166-1174. https://doi.org/10.1002/jcb.22345
Pucci, Bruna ; Indelicato, Manuela ; Paradisi, Valentina ; Reali, Valentina ; Pellegrini, Laura ; Aventaggiato, Michele ; Karpinich, Natalie O. ; Fini, Massimo ; Russo, Matteo A. ; Farber, John L. ; Tafani, Marco. / ERK-1 MAP kinase prevents TNF-induced apoptosis through bad phosphorylation and inhibition of bax translocation in HeLa cells. In: Journal of Cellular Biochemistry. 2009 ; Vol. 108, No. 5. pp. 1166-1174.
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