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


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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Figure 1: Defective proinflammatory cytokine production in Ubc13-deficient bone marrow macrophages.
Figure 2: Impaired B cell development in Ubc13fl/flCd19-Cre mice.
Figure 3: Ubc13 is required for B cell activation and in vivo immune responses.
Figure 4: NF-κB activation in Ubc13-deficient cells.
Figure 5: Impaired MAP kinase activation in Ubc13-deficient cells.
Figure 6: Ubc13-dependent ubiquitination of IKKγ in IL-1- and TLR-induced Jnk activation.


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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

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.

Correspondence to Shizuo Akira.

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The authors declare no competing financial interests.

Supplementary information

Supplementary Fig. 1

Generation of conditional Ubc13fl/fl mice (PDF 217 kb)

Supplementary Fig. 2

CD3+ T cells in spleens of Ubc13fl/flCd19-Cre mice. (PDF 123 kb)

Supplementary Fig. 3

NF-κB complexes and NF-κBp100 processing in Ubc13-deficient B cells. (PDF 317 kb)

Supplementary Fig. 4

Defective activation of MAP kinases and increased IRAK-1 ubiquitination in IKKγ-deficient cell lines (PDF 225 kb)

Supplementary Table 1

Genotype analysis of offspring from UBC13 heterozygous intercrosses. (PDF 71 kb)

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