A Novel Interaction between the Juxtamembrane Region of the p55 Tumor Necrosis Factor Receptor and Phosphatidylinositol-4-phosphate 5-Kinase

Alexander M Castellino,Gregory J Parker,Igor V Boronenkov,Richard A Anderson,Moses V Chao

doi:10.1074/jbc.272.9.5861

Alexander M Castellino, Gregory J Parker + Show 3 more

Open Access

https://doi.org/10.1074/jbc.272.9.5861

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Abstract

Tumor necrosis factor-alpha (TNF-alpha) binding to its receptors leads to a diversity of biological responses. The actions of TNF are the result of the interaction of cytoplasmic proteins that bind directly to the intracellular domains of the two TNF receptors, p55 and p75. Here we report a novel interaction between the juxtamembrane region of the p55 TNF receptor and a newly discovered 47-kDa isoform of phosphatidylinositol-4-phosphate 5-kinase (PIP5K), a member of the enzyme family that generates the key signaling messenger, phosphatidylinositol 4,5-bisphosphate. The interaction was found to be specific for the p55 TNF receptor and was not observed with the p75 TNF receptor, the Fas antigen, or the p75 neurotrophin receptor, which are other members of the TNF receptor superfamily. In vitro experiments using recombinant fusion proteins verify the authenticity of the interaction between the p55 receptor and PIP5KIIbeta, a new isoform of PIP5K, but not the previously identified 53-kDa PIP5KIIalpha. Treatment of HeLa cells with TNF-alpha resulted in an increased PIP5K activity. These results indicate that phosphatidylinositol turnover may be linked to stimulation of the p55 TNF receptor and suggest that a subset of TNF responses may result from the direct association of PIP5KIIbeta with the p55 TNF receptor.

Highlights

Tumor necrosis factor-␣ (TNF-␣)1 initiates its proliferative, differentiative, or cytotoxic actions on mammalian cells by binding to two transmembrane molecules, the p55 and p75 TNF receptors [1,2,3]
While overexpression of TRADD or FADD in heterologous cells leads to cell death [7, 11], deletions or mutations in the death domains abolish the ability of these molecules to participate in the initiation of apoptosis [6, 12]
The connection of the Fas antigen and the p55 TNF receptor with a member of the interleukin converting enzyme protease family provides a mechanism for how cell death signals are initiated by ligand-receptor interactions

Summary

Introduction

Tumor necrosis factor-␣ (TNF-␣)1 initiates its proliferative, differentiative, or cytotoxic actions on mammalian cells by binding to two transmembrane molecules, the p55 and p75 TNF receptors [1,2,3]. We report the interaction of the juxtamembrane region of the p55 TNF receptor with phosphatidylinositol-4-phosphate 5-kinase (PIP5K), an enzyme that produces phosphatidylinositol 4,5-bisphosphate (PIP2). To identify cellular proteins that bound to the juxtamembrane domain of the p55 TNF receptor, a recombinant LexA fusion protein was generated that contained amino acids 204 –280 (Y1), beginning at the end of the transmembrane domain and extending to the middle of the cytoplasmic domain.

Results

Conclusion