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Key function for the Ubc13 E2 ubiquitin-conjugating enzyme in immune receptor signaling

Nature Immunology volume 7, pages 962970 (2006) | Download Citation

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Abstract

The Ubc13 E2 ubiquitin-conjugating enzyme is key in the process of 'tagging' target proteins with lysine 63–linked polyubiquitin chains, which are essential for the transmission of immune receptor signals culminating in activation of the transcription factor NF-κB. Here we demonstrate that conditional ablation of Ubc13 resulted in defective B cell development and in impaired B cell and macrophage activation. In response to all tested stimuli except tumor necrosis factor, Ubc13-deficient cells showed almost normal NF-κB activation but considerably impaired activation of mitogen-activated protein kinase. Ubc13-induced activation of mitogen-activated protein kinase required, at least in part, ubiquitination of the adaptor protein IKKγ. These results show that Ubc13 is key in the mammalian immune response.

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Acknowledgements

We thank T. Kitamura (The University of Tokyo, Tokyo, Japan) for Plat-E packaging cell lines; D.T. Golenbock (University of Massachusetts Medical School, Worcester, Massachusetts) for the NF-κB-dependent ELAM1 reporter plasmid; J.L. Pomerantz (The Johns Hopkins University School of Medicine, Baltimore, Maryland) for Card11 expression vectors; J. Inoue (The University of Tokyo, Tokyo, Japan) for the pFastBacHTa-TRAF6 vector; G. Courtois (Hôpital Saint-Louis, Paris, France) for the IKKγ-deficient 1.3E2 cell line; M. Pasparakis (European Molecular Biology Laboratory, Rome, Italy) for MEFs from IKKγ-deficient mice; R.C. Rickert (The Burnham Institute, La Jolla, California) for Cd19-Cre mice; I. Förster (University of Munich, Munich, Germany) for LyzsM-Cre mice; H. Hemmi, T. Yasui and T. Matsunaga for discussions; M. Hashimoto for secretarial assistance; and N. Okita, N. Iwami, N. Fukuda and M. Morita for technical assistance. Supported by Special Coordination Funds; the Ministry of Education, Culture, Sports, Science and Technology; Research Fellowships of the Japan Society for the Promotion of Science for Young Scientists; The Uehara Memorial Foundation; The Naito Foundation and The Junior Research Associate from RIKEN; and Exploratory Research for Advanced Technology, Japan Science and Technology Agency.

Author information

Affiliations

  1. Department of Host Defense, Research Institute for Microbial Diseases, Osaka University, Osaka 565-0871, Japan.

    • Masahiro Yamamoto
    • , Hideki Sanjo
    • , Satoshi Uematsu
    • , Tatsuya Saitoh
    • , Osamu Takeuchi
    •  & Shizuo Akira
  2. Department of Molecular Virology, Research Institute for Microbial Diseases, Osaka University, Osaka 565-0871, Japan.

    • Toru Okamoto
    •  & Yoshiharu Matsuura
  3. Department of Embryonic and Genetic Engineering, Medical Institute of Bioregulation, Kyushu University, Fukuoka 812-8582, Japan.

    • Kiyoshi Takeda
  4. ERATO, Japan Science and Technology Corporation, Osaka 565-0871, Japan.

    • Shintaro Sato
    • , Ken J Ishii
    • , Taro Kawai
    • , Osamu Takeuchi
    •  & Shizuo Akira
  5. Department of Molecular Virology, Graduate School of Medicine, Tokyo Medical and Dental University, Tokyo 113-8519, Japan.

    • Tatsuya Saitoh
    • , Naoki Yamamoto
    •  & Shoji Yamaoka
  6. Division of Pathogenic Biochemistry, Institute of Natural Medicine, 21st Century Center of Excellence Program, Toyama Medical and Pharmaceutical University, Toyama 930-0194, Japan.

    • Hiroaki Sakurai

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Contributions

All authors contributed to data analysis, experimental design, critical discussions and manuscript preparation; M.Y did all experimental studies; T.O. and Y.M. purified recombinant TRAF6; K.T. generated Ubc13-deficient mice; S.S. and S.U. prepared whole-cell extracts; T.S., N.Y. and S.Y. designed retroviral vectors; H.S. prepared antibodies; K.J.I., T.K. and O.T. played a pivotal role in discussions; and S.A. supervised all work.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Shizuo Akira.

Supplementary information

PDF files

  1. 1.

    Supplementary Fig. 1

    Generation of conditional Ubc13fl/fl mice

  2. 2.

    Supplementary Fig. 2

    CD3+ T cells in spleens of Ubc13fl/flCd19-Cre mice.

  3. 3.

    Supplementary Fig. 3

    NF-κB complexes and NF-κBp100 processing in Ubc13-deficient B cells.

  4. 4.

    Supplementary Fig. 4

    Defective activation of MAP kinases and increased IRAK-1 ubiquitination in IKKγ-deficient cell lines

  5. 5.

    Supplementary Table 1

    Genotype analysis of offspring from UBC13 heterozygous intercrosses.

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DOI

https://doi.org/10.1038/ni1367

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