Pellino 3b Negatively Regulates Interleukin-1-induced TAK1-dependent NFκB Activation

Hui Xiao,Wen Qian,Kirk Staschke,Youcun Qian,Grace Cui,Li Deng,Mariam Ehsani,Xiliang Wang,Yue-Wei Qian,Zhijian J Chen,Raymond Gilmour,Zhengfan Jiang,Xiaoxia Li

doi:10.1074/jbc.m706931200

Abstract

Pellino 3b Negatively Regulates Interleukin-1-induced TAK1-dependent NFκB Activation

Highlights

14654 JOURNAL OF BIOLOGICAL CHEMISTRY ing to hyperphosphorylation of IL-1 receptor-associated kinase (IRAK) [10], which creates an interface for its interaction with adapter Pellino 1 [11]
To examine whether IL-1-induced polyubiquitination of IRAK is linked via Lys-63 or Lys-48, HA-tagged wild type and a series of mutants of ubiquitin were transfected into 293 cells
Following in vitro ubiquitination assay, 20 ␮l of Laemmli buffer was added into 10 ␮l of reaction. 15 ␮l of the reaction mixture was analyzed by Western blotting (WB) with anti-ubiquitin, and the other half was visualized by Coomassie Blue staining after resolving by 10% SDS-PAGE

Summary

Introduction

14654 JOURNAL OF BIOLOGICAL CHEMISTRY ing to hyperphosphorylation of IRAK [10], which creates an interface for its interaction with adapter Pellino 1 [11]. Pellino 3b failed to inhibit found that IL-1-induced TAK1 modification and I␬B␣ degra- TNF␣-induced I␬B␣ phosphorylation, I␬B␣ degradation, or dation were reduced in Pellino 2 knockdown cells, suggesting JNK activation (Fig. 8E), indicating the specific role of Pellino the importance of Pellino 2 in TAK1-dependent NF␬B activa- 3b in regulating IL-1 signaling.

Results

Conclusion