HTLV-1 Tax Induces Formation of the Active Macromolecular IKK Complex by Generating Lys63- and Met1-Linked Hybrid Polyubiquitin Chains

PLoS Pathogens

Satoshi Inoue¹  Hiroyasu Nakano²  Tsukasa Seya³  Hiroyuki Oshiumi³  Kazuhiro Iwai⁴  Fuminori Tokunaga⁵  Eiji Goto⁵  Ginga Komatsu⁶  Yuri Shibata⁶  Jun-ichiro Inoue⁶  Yuetsu Tanaka⁷  Yasushi Saeki⁸  Keiji Tanaka⁸  Hirotaka Takahashi⁹  Tatsuya Sawasaki⁹  Jin Gohda^1,10 

[1] Department of Anti-Aging Medicine, Graduate School of Medicine, The University of Tokyo, Tokyo, Japan;Department of Biochemistry, Toho University School of Medicine, Tokyo, Japan;Department of Microbiology and Immunology, Graduate School of Medicine, Hokkaido University, Sapporo, Japan;Department of Molecular and Cellular Physiology, Graduate School of Medicine, Kyoto University, Kyoto, Japan;Department of Pathobiochemistry, Graduate School of Medicine, Osaka City University, Osaka, Japan;Division of Cellular and Molecular Biology, Department of Cancer Biology, The Institute of Medical Science, The University of Tokyo, Tokyo, Japan;Division of Immunology, Faculty of Medicine, University of the Ryukyus, Okinawa, Japan;Laboratory of Protein Metabolism, Tokyo Metropolitan Institute of Medical Science, Tokyo, Japan;Proteo-Science Center, Ehime University, Ehime, Japan;Research Center for Asian Infectious Diseases, The Institute of Medical Science, The University of Tokyo, Tokyo, Japan

关键词: Immunoprecipitation;    Immunoblotting;    Transcription factors;    Recombination reactions;    Phosphorylation;    T cells;    HTLV-1;    Ligases;   

DOI  :  10.1371/journal.ppat.1006162

学科分类：生物科学（综合）

来源: Public Library of Science

【 摘 要 】

The Tax protein of human T-cell leukemia virus type 1 (HTLV-1) is crucial for the development of adult T-cell leukemia (ATL), a highly malignant CD4+ T cell neoplasm. Among the multiple aberrant Tax-induced effects on cellular processes, persistent activation of transcription factor NF-κB, which is activated only transiently upon physiological stimulation, is essential for leukemogenesis. We and others have shown that Tax induces activation of the IκB kinase (IKK) complex, which is a critical step in NF-κB activation, by generating Lys63-linked polyubiquitin chains. However, the molecular mechanism underlying Tax-induced IKK activation is controversial and not fully understood. Here, we demonstrate that Tax recruits linear (Met1-linked) ubiquitin chain assembly complex (LUBAC) to the IKK complex and that Tax fails to induce IKK activation in cells that lack LUBAC activity. Mass spectrometric analyses revealed that both Lys63-linked and Met1-linked polyubiquitin chains are associated with the IKK complex. Furthermore, treatment of the IKK-associated polyubiquitin chains with Met1-linked-chain-specific deubiquitinase (OTULIN) resulted in the reduction of high molecular weight polyubiquitin chains and the generation of short Lys63-linked ubiquitin chains, indicating that Tax can induce the generation of Lys63- and Met1-linked hybrid polyubiquitin chains. We also demonstrate that Tax induces formation of the active macromolecular IKK complex and that the blocking of Tax-induced polyubiquitin chain synthesis inhibited formation of the macromolecular complex. Taken together, these results lead us to propose a novel model in which the hybrid-chain-dependent oligomerization of the IKK complex triggered by Tax leads to trans-autophosphorylation-mediated IKK activation.

【 授权许可】

CC BY