Molecular bases for HOIPINs-mediated inhibition of LUBAC and innate immune responses

Daisuke Oikawa; Yusuke Sato; Fumiaki Ohtake; Keidai Komakura; Kazuki Hanada; Koji Sugawara; Seigo Terawaki; Yukari Mizukami; Hoang T Phuong; Kiyosei Iio; Shingo Obika; Masaya Fukushi; Takashi Irie; Daisuke Tsuruta; Shinji Sakamoto; Keiji Tanaka; Yasushi Saeki; Shuya Fukai; Fuminori Tokunaga

doi:10.1038/s42003-020-0882-8

Molecular bases for HOIPINs-mediated inhibition of LUBAC and innate immune responses

Commun Biol. 2020 Apr 3;3(1):163. doi: 10.1038/s42003-020-0882-8.

Authors

Daisuke Oikawa¹, Yusuke Sato^{2

3

4}, Fumiaki Ohtake^{5

6}, Keidai Komakura^{1

7}, Kazuki Hanada⁸, Koji Sugawara⁷, Seigo Terawaki¹, Yukari Mizukami⁷, Hoang T Phuong⁷, Kiyosei Iio⁹, Shingo Obika⁹, Masaya Fukushi¹⁰, Takashi Irie¹⁰, Daisuke Tsuruta⁷, Shinji Sakamoto⁸, Keiji Tanaka⁵, Yasushi Saeki⁵, Shuya Fukai^{2

3}, Fuminori Tokunaga¹¹

Affiliations

¹ Department of Pathobiochemistry, Graduate School of Medicine, Osaka City University, Osaka, Japan.
² Institute for Quantitative Biosciences, The University of Tokyo, Tokyo, Japan.
³ Synchrotron Radiation Research Organization, The University of Tokyo, Tokyo, Japan.
⁴ Center for Research on Green Sustainable Chemistry, Tottori University, Tottori, Japan.
⁵ Laboratory of Protein Metabolism, Tokyo Metropolitan Institute of Medical Science, Tokyo, Japan.
⁶ Institute for Advanced Life Sciences, Hoshi University, Tokyo, Japan.
⁷ Department of Dermatology, Graduate School of Medicine, Osaka City University, Osaka, Japan.
⁸ Pharmaceutical Frontier Research Laboratories, Central Pharmaceutical Research Institute, JT Inc., Kanagawa, Japan.
⁹ Chemical Research Laboratories, Central Pharmaceutical Research Institute, JT Inc., Osaka, Japan.
¹⁰ Department of Virology, Graduate School of Biomedical and Health Sciences, Hiroshima University, Hiroshima, Japan.
¹¹ Department of Pathobiochemistry, Graduate School of Medicine, Osaka City University, Osaka, Japan. [email protected].

Abstract

The NF-κB and interferon antiviral signaling pathways play pivotal roles in inflammatory and innate immune responses. The LUBAC ubiquitin ligase complex, composed of the HOIP, HOIL-1L, and SHARPIN subunits, activates the canonical NF-κB pathway through Met1-linked linear ubiquitination. We identified small-molecule chemical inhibitors of LUBAC, HOIPIN-1 and HOIPIN-8. Here we show that HOIPINs down-regulate not only the proinflammatory cytokine-induced canonical NF-κB pathway, but also various pathogen-associated molecular pattern-induced antiviral pathways. Structural analyses indicated that HOIPINs inhibit the RING-HECT-hybrid reaction in HOIP by modifying the active Cys885, and residues in the C-terminal LDD domain, such as Arg935 and Asp936, facilitate the binding of HOIPINs to LUBAC. HOIPINs effectively induce cell death in activated B cell-like diffuse large B cell lymphoma cells, and alleviate imiquimod-induced psoriasis in model mice. These results reveal the molecular and cellular bases of LUBAC inhibition by HOIPINs, and demonstrate their potential therapeutic uses.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

A549 Cells
Animals
Anti-Inflammatory Agents / chemistry
Anti-Inflammatory Agents / pharmacology*
Antineoplastic Agents / chemistry
Antineoplastic Agents / pharmacology*
Apoptosis / drug effects
Disease Models, Animal
Enzyme Inhibitors / chemistry
Enzyme Inhibitors / pharmacology*
Female
HEK293 Cells
HeLa Cells
Humans
Imiquimod
Immunity, Innate / drug effects*
Inflammation Mediators / metabolism
Intracellular Signaling Peptides and Proteins / genetics
Intracellular Signaling Peptides and Proteins / metabolism
Jurkat Cells
Lymphoma, Large B-Cell, Diffuse / drug therapy*
Lymphoma, Large B-Cell, Diffuse / immunology
Lymphoma, Large B-Cell, Diffuse / metabolism
Lymphoma, Large B-Cell, Diffuse / pathology
Mice
Mice, Inbred BALB C
Molecular Structure
Psoriasis / chemically induced
Psoriasis / immunology
Psoriasis / metabolism
Psoriasis / prevention & control*
Signal Transduction
Structure-Activity Relationship
Transcription Factors / genetics
Transcription Factors / metabolism
Ubiquitin-Protein Ligases / antagonists & inhibitors*
Ubiquitin-Protein Ligases / genetics
Ubiquitin-Protein Ligases / metabolism
Ubiquitins / genetics
Ubiquitins / metabolism

Substances

Anti-Inflammatory Agents
Antineoplastic Agents
Enzyme Inhibitors
Inflammation Mediators
Intracellular Signaling Peptides and Proteins
SHARPIN protein, human
Sipl1 protein, mouse
Transcription Factors
Ubiquitins
RBCK1 protein, human
RNF31 protein, human
Rbck1 protein, mouse
Rnf31 protein, mouse
Ubiquitin-Protein Ligases
Imiquimod