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PDLIM2-mediated termination of transcription factor NF-κB activation by intranuclear sequestration and degradation of the p65 subunit

Nature Immunology volume 8, pages 584–591 (2007)Cite this article

Abstract

Activation of transcription factor NF-κB in the innate immune system is tightly regulated to prevent excessive inflammatory responses. How NF-κB activation is terminated, however, is not fully understood. Here we report that PDLIM2 negatively regulated NF-κB activity, acting as a nuclear ubiquitin E3 ligase targeting the p65 subunit of NF-κB. PDLIM2 bound to p65 and promoted p65 polyubiquitination. In addition, PDLIM2 targeted p65 to discrete intranuclear compartments where polyubiquitinated p65 was degraded by the proteasome. PDLIM2 deficiency resulted in larger amounts of nuclear p65, defective p65 ubiquitination and augmented production of proinflammatory cytokines in response to innate stimuli. Our findings delineate a pathway by which PDLIM2 terminates NF-κB activation through intranuclear sequestration and subsequent degradation.

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Figure 1: Ubiquitin- and proteasome-dependent degradation of endogenous p65.
Figure 2: PDLIM2 binds to and polyubiquitinates p65 and negatively regulates NF-κB-mediated transcription.
Figure 3: PDLIM2 targets p65 to insoluble nuclear compartments, where p65 is degraded by the proteasome.
Figure 4: PDLIM2 promotes the intranuclear translocation of nuclear p65 into PML nuclear bodies by means of its PDZ domain.
Figure 5: Enhanced p65 activation in Pdlim2−/− mice.

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Acknowledgements

We thank K. Hoshino, T. Sugiyama, M. Saito for discussions; N. Okita, N. Iwami, Y. Fukuda and E. Haga for technical assistance; S. Haraguchi for secretarial assistance; A. Furuno for help with confocal microscopy; and R. Triendl, L. Leclercq and P. Burrows for critical review of the manuscript. The plasmid pCII-CMV-MCS-IRES-Venus was provided by H. Miyoshi (University of Tsukuba) and A. Miyawaki (RIKEN Brain Science Institute), and the ELAM-1 luciferase reporter construct was provided by D. Golenbock (University of Massachusetts Medical School). Supported by Scientific Research of MEXT (Ministry of Education, Culture, Sports. Science and Technology) and the Japan Society for the Promotion of Science (T.T. and T.K.), the Mochida Memorial Foundation for Medical and Pharmaceutical Research (T.T. and T.K.), the RIKEN Strategic Research Program (T.T. and T.K.), the Uehara Memorial Foundation (T.K.), the Sumitomo Foundation (T.T.), the National Institutes of Health (AI506296 to M.J.G.) and the G. Harold and Leila Y. Mathers Charitable Foundation (M.J.G.).

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Authors and Affiliations

  1. Laboratory for Host Defense, RIKEN Research Center for Allergy and Immunology, Yokohama, 230-0045, Kanagawa, Japan

    Takashi Tanaka & Tsuneyasu Kaisho

  2. Department of Immunology and Infectious Diseases, Harvard School of Public Health, Boston, 02115, Massachusetts, USA

    Michael J Grusby

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  1. Takashi Tanaka
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T.T. contributed to general experiments and design with the direction and supervision of T.K.; M.J.G. contributed to discussions and manuscript criticism.

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Correspondence to Tsuneyasu Kaisho.

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Supplementary information

Supplementary Fig. 1

Interaction of PDLIM2 mutants lacking LIM or PDZ domains with endogenous p65 is comparable to that of wild-type PDLIM2. (PDF 67 kb)

Supplementary Fig. 2

PDZ domain of PDLIM2 is not required for p65 ubiquitination. (PDF 47 kb)

Supplementary Fig. 3

Enhanced proinflammatory cytokine production in Pdlim2−/− cells in the absence of IFN-γ. (PDF 41 kb)

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Tanaka, T., Grusby, M. & Kaisho, T. PDLIM2-mediated termination of transcription factor NF-κB activation by intranuclear sequestration and degradation of the p65 subunit. Nat Immunol 8, 584–591 (2007). https://doi.org/10.1038/ni1464

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