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Intracellular action of the cytokine MIF to modulate AP-1 activity and the cell cycle through Jab1

Abstract

Cytokines are multifunctional mediators that classically modulate immune activity by receptor-mediated pathways. Macrophage migration inhibitory factor (MIF) is a cytokine that has a critical role in several inflammatory conditions1,2,3 but that also has endocrine4,5 and enzymatic functions6,7. The molecular targets of MIF action have so far remained unclear. Here we show that MIF specifically interacts with an intracellular protein, Jab1, which is a coactivator of AP-1 transcription8,9 that also promotes degradation of the cyclin-dependent kinase inhibitor p27Kip1 (ref. 10). MIF colocalizes with Jab1 in the cytosol, and both endogenous and exogenously added MIF following endocytosis bind Jab1. MIF inhibits Jab1- and stimulus-enhanced AP-1 activity, but does not interfere with the induction of the transcription factor NFκB. Jab1 activates c-Jun amino-terminal kinase (JNK) activity and enhances endogenous phospho-c-Jun levels, and MIF inhibits these effects. MIF also antagonizes Jab1-dependent cell-cycle regulation by increasing p27Kip1 expression through stabilization of p27Kip1 protein. Consequently, Jab1-mediated rescue of fibroblasts from growth arrest is blocked by MIF. Amino acids 50–65 and Cys 60 of MIF are important for Jab1 binding and modulation. We conclude that MIF may act broadly to negatively regulate Jab1-controlled pathways and that the MIF–Jab1 interaction may provide a molecular basis for key activities of MIF.

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Figure 1: MIF specifically interacts with Jab1.
Figure 2: MIF is targeted to the cytosol and colocalizes with Jab1.
Figure 3: MIF inhibits stimulated AP-1 activity.
Figure 4: Jab1 enhances JNK activity and phospho-c-Jun levels and MIF inhibits these effects.
Figure 5: MIF stabilizes p27Kip1 protein and inhibits fibroblast growth in a Jab1-dependent manner.

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Acknowledgements

We thank S. Schlenker and E. Wagner for experimental assistance; P. Angel for the pBAT-c-Jun and pRSV-c-Jun constructs; and R. Bucala for communicating unpublished data on MIF/Erk induction. We thank A. Billich/ Novartis Research Center, Vienna, for testing the effect of anti-MIF antibodies on fibroblast growth. We are grateful to K. Pfizenmaier, F. Vitzhum, and K. Sohn for helpful discussions and critical reading of the manuscript. This work was supported by a grant from the Fraunhofer IGB.

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Correspondence to Jürgen Bernhagen.

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Kleemann, R., Hausser, A., Geiger, G. et al. Intracellular action of the cytokine MIF to modulate AP-1 activity and the cell cycle through Jab1. Nature 408, 211–216 (2000). https://doi.org/10.1038/35041591

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