β-actin is required for mitochondria clustering and ROS generation in TNF-induced, caspase-independent cell death
Open Access
- 15 September 2004
- journal article
- Published by The Company of Biologists in Journal of Cell Science
- Vol. 117 (20) , 4673-4680
- https://doi.org/10.1242/jcs.01339
Abstract
Tumor necrosis factor (TNF)-α induces caspase-independent cell death in the fibrosarcoma cell line L929. This cell death has a necrotic phenotype and is dependent on production of reactive oxygen species (ROS) in the mitochondria. To identify genes involved in this TNF-induced, ROS-dependent cell death pathway, we utilized retrovirus insertion-mediated random mutagenesis to generate TNF-resistant L929 cell lines and we subsequently identified genes whose mutations are responsible for the TNF-resistant phenotype. In one such resistant line, β-actin was disrupted by viral insertion, and subsequent reconstitution of β-actin expression levels in the mutant line Actinmut restored its sensitivity to TNF. Resistance to TNF in Actinmut cells is signal specific since the sensitivity to other death stimuli is either unchanged or even increased. Comparable NF-κB activation and p38 phosphorylation in TNF-treated wild-type and Actinmut cells also indicates that reduced expression of actin only selectively blocked some of the TNF-induced cellular changes. Actin cleavage involved in apoptosis does not occur in TNF-treated L929 cell death, as in HeLa cells. Consistent over-expression of a caspase-cleaved product, a 15 kDa actin fragment, had no effect on TNF-induced necrosis of L929 cell. By contrast, TNF-induced mitochondria clustering and ROS production were dramatically reduced in Actinmut cells, indicating that actin-deficiency-mediated TNF resistance is most likely due to impaired mitochondrial responses to TNF stimulation. Our findings suggest that a full complement of actin is required for transduction of a cell death signal to mitochondria in TNF-treated L929 cells.Keywords
This publication has 53 references indexed in Scilit:
- Inhibition of actin polymerization enhances commitment to and execution of apoptosis induced by withdrawal of trophic supportJournal of Cellular Biochemistry, 2003
- Actin Cytoskeleton Is Required for Early Apoptosis Signaling Induced by Anti-Fas Antibody but Not Fas Ligand in Murine B Lymphoma A20 CellsBiochemical and Biophysical Research Communications, 2002
- The p65/RelA Subunit of NF-κB Interacts with Actin-Containing StructuresExperimental Cell Research, 2000
- Biochemical Pathways of Caspase Activation During ApoptosisAnnual Review of Cell and Developmental Biology, 1999
- Involvement of MACH, a Novel MORT1/FADD-Interacting Protease, in Fas/APO-1- and TNF Receptor–Induced Cell DeathCell, 1996
- Molecular mechanisms of tumor necrosis factor‐induced cytotoxicityFEBS Letters, 1994
- Butylated hydroxyanisole specifically inhibits tumor necrosis factor-induced cytotoxicity and growth enhancementCytokine, 1992
- TUMOR NECROSIS, CACHEXIA, SHOCK, AND INFLAMMATION: A COMMON MEDIATORAnnual Review of Biochemistry, 1988
- Cachectin and tumour necrosis factor as two sides of the same biological coinNature, 1986
- An endotoxin-induced serum factor that causes necrosis of tumors.Proceedings of the National Academy of Sciences, 1975