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Positive regulation of immune cell function and inflammatory responses by phosphatase PAC-1

Nature Immunology volume 7, pages 274–283 (2006)Cite this article

Abstract

Mitogen-activated protein kinases facilitate many cellular processes and are essential for immune cell function. Their activity is controlled by kinases and dual-specificity phosphatases. A comprehensive microarray analysis of human leukocytes identified DUSP2 (encoding the phosphatase PAC-1) as one of the most highly induced transcripts in activated immune cells. We generated Dusp2−/− mice and found considerably reduced inflammatory responses in the 'K/BxN' model of rheumatoid arthritis. PAC-1 deficiency led to increased activity of Jun kinase (Jnk) but unexpected impairment of the activity of extracellular signal–regulated kinase (Erk) and the kinase p38, reduced activity of the transcription factor Elk1 and a complex of mobilized transcription factor NFAT and the AP-1 transcription factor and decreased effector immune cell function. Thus, PAC-1 is a key positive regulator of inflammatory cell signaling and effector functions, mediated through Jnk and Erk mitogen-activated protein kinase crosstalk.

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Figure 1: DUSP2 is the most highly induced DUSP transcript in activated human leukocytes, and PAC-1 protein associates with inflammatory tissue.
Figure 2: Targeted deletion of mouse Dusp2 results in protection of Dusp2−/− mice in the K/BxN model of rheumatoid arthritis.
Figure 3: Dusp2−/− macrophages and mast cells are compromised in their production of proinflammatory mediators.
Figure 4: Re-expression of Dusp2 enhances or restores cytokine production in a phosphatase activity–dependent way.
Figure 5: Targeted deletion of Dusp2 results in decreased Erk and p38 kinase activity and increased phosphorylated Jnk.
Figure 6: Reduced transcriptional Elk1 and NFAT–AP-1 activity in Dusp2−/− BMMCs.
Figure 7: Inhibition of Jnk can 'rescue' the reduced phosphorylated Erk in Dusp2−/− BMMCs and increase Elk1 transcriptional activity.

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Acknowledgements

We thank R. Daly for suggestions; D. Mathis (Harvard Medical School, Boston, Massachusetts) and C. Benoist for supplying KRN mice; R. Newton and C. Waltzinger for flow cytometry assistance; J. Thatcher and D. Zhara for PAC-1 antibodies; W. Kaplan for statistical assistance with mast cell GeneChip data; and T. Kitamura for Plat-E cells, pMx vector and technical advice on retroviral transductions. C.R.M. and S.D.G. share senior authorship. Supported by National Health and Medical Research Council of Australia and the Cooperative Research Center for Asthma.

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Authors and Affiliations

  1. Garvan Institute of Medical Research, Sydney, 2010, New South Wales, Australia

    Kate L Jeffrey, Tilman Brummer, Michael S Rolph, Sue M Liu, Nuria A Callejas, Fabienne Mackay, Shane Grey & Charles R Mackay

  2. Cooperative Research Centre for Asthma, Darlinghurst, Sydney, 2010, New South Wales, Australia

    Kate L Jeffrey, Michael S Rolph, Sue M Liu & Charles R Mackay

  3. Walter and Eliza Hall Institute, Parkville, Melbourne, 3050, Victoria, Australia

    Raelene J Grumont & Steve D Gerondakis

  4. Serono Pharmaceutical Research Institute, Serono International, Geneva, 1228, Switzerland

    Corine Gillieron, Montserrat Camps & Christian Rommel

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Correspondence to Steve D Gerondakis or Charles R Mackay.

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Supplementary information

Supplementary Fig. 1

PAC-1 is hematopoietic cell-specific in its expression. (PDF 25 kb)

Supplementary Fig. 2

Selected PAC-1 monoclonal antibodies show good selectivity towards PAC-1 over other related DUSPs and other phosphatases. (PDF 65 kb)

Supplementary Fig. 3

Genechip expression profiling of DUSPs in mouse leukocytes, qPCR of pac-1 mRNA and immunoblot of PAC-1 in primary immune cells. (PDF 72 kb)

Supplementary Fig. 4

Targeted deletion of mouse Pac1 gene results in protection of Pac1−/− mice in the K/BxN model of rheumatoid arthritis. (PDF 185 kb)

Supplementary Fig. 5

Pac1+/+ and Pac1−/− BMMCs have comparable physical and functional properties. (PDF 40 kb)

Supplementary Fig. 6

Further analysis of the differential expression of signal transducers and transcription factors between wild-type and PAC-1 deficient activated macrophages and mast cells. (PDF 192 kb)

Supplementary Fig. 7

Elk-1 activity is dependent on ERK activity and inhibition of both ERK and p38 can eliminate TNF and IL-4 production in BMMCs. (PDF 24 kb)

Supplementary Methods (PDF 45 kb)

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Jeffrey, K., Brummer, T., Rolph, M. et al. Positive regulation of immune cell function and inflammatory responses by phosphatase PAC-1. Nat Immunol 7, 274–283 (2006). https://doi.org/10.1038/ni1310

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