Document Type
Article
Publication Date
4-3-2020
Abstract
The necrosome is a protein complex required for signaling in cells that results in necroptosis, which is also dependent on tumor necrosis factor receptor (TNF-R) signaling. TNFα promotes necroptosis, and its expression is facilitated by mitogen-activated protein (MAP) kinase-activated protein kinase 2 (MK2) but is inhibited by the RNA-binding protein tristetraprolin (TTP, encoded by the Zfp36 gene). We have stimulated murine macrophages from WT, MyD88 -/-, Trif -/-, MyD88 -/- Trif -/-, MK2 -/-, and Zfp36 -/- mice with graded doses of lipopolysaccharide (LPS) and various inhibitors to evaluate the role of various genes in Toll-like receptor 4 (TLR4)-induced necroptosis. Necrosome signaling, cytokine production, and cell death were evaluated by immunoblotting, ELISA, and cell death assays, respectively. We observed that during TLR4 signaling, necrosome activation is mediated through the adaptor proteins MyD88 and TRIF, and this is inhibited by MK2. In the absence of MK2-mediated necrosome activation, lipopolysaccharide-induced TNFα expression was drastically reduced, but MK2-deficient cells became highly sensitive to necroptosis even at low TNFα levels. In contrast, during tonic TLR4 signaling, WT cells did not undergo necroptosis, even when MK2 was disabled. Of note, necroptosis occurred only in the absence of TTP and was mediated by the expression of TNFα and activation of JUN N-terminal kinase (JNK). These results reveal that TTP plays an important role in inhibiting TNFα/JNK-induced necrosome signaling and resultant cytotoxicity.
Recommended Citation
Ariana, Ardeshir; Alturki, Norah A; Hajjar, Stephanie; Stumpo, Deborah J; Tiedje, Christopher; Alnemri, Emad S; Gaestel, Matthias; Blackshear, Perry J; and Sad, Subash, "Tristetraprolin regulates necroptosis during tonic Toll-like receptor 4 (TLR4) signaling in murine macrophages." (2020). Department of Biochemistry and Molecular Biology Faculty Papers. Paper 166.
https://jdc.jefferson.edu/bmpfp/166
PubMed ID
32094226
Language
English
Comments
This research was originally published in the Journal of Biological Chemistry. Ariana, Ardeshir; Alturki, Norah A; Hajjar, Stephanie; Stumpo, Deborah J; Tiedje, Christopher; Alnemri, Emad S; Gaestel, Matthias; Blackshear, Perry J; and Sad, Subash, "Tristetraprolin regulates necroptosis during tonic Toll-like receptor 4 (TLR4) signaling in murine macrophages."J Biol Chem. 2020; 284(14):661-4672. the Author(s)."
The published article can be found at https://doi.org/10.1074/jbc.RA119.011633.