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Interaction between TAK1–TAB1–TAB2 and RCAN1–calcineurin defines a signalling nodal control point

Nature Cell Biology volume 11, pages 154–161 (2009)Cite this article

Abstract

The calcium-activated protein phosphatase calcineurin is controlled by regulator of calcineurin (RCAN) in organisms ranging from yeast to mammals. Here we performed a yeast two-hybrid screen with RCAN1 as bait, identifying TAK1 binding protein 2 (TAB2) as an interacting partner. TAB2 interacted directly with RCAN1 in vitro and in vivo, recruiting TAK1, TAB1 and calcineurin, forming a macromolecular signalling complex. Overexpression of TAK1 and TAB1, or active TAK1ΔN, promoted direct phosphorylation of RCAN1 in vitro and in vivo. TAK1 phosphorylated RCAN1 at Ser 94 and Ser 136, converting RCAN1 from an inhibitor to a facilitator of calcineurin–NFAT signalling, and enhancing NFATc1 nuclear translocation, NFAT transcriptional activation and the hypertrophic growth of cultured cardiomyocytes. The TAK1–TAB1–TAB2 and the calcineurin–NFAT signalling modules did not interact in Rcan1/2- or Tab2-deficient mouse embryonic fibroblast (MEF) cultures. Calcineurin activation also dephosphorylated and inhibited TAK1 and TAB1, an effect that was absent in Rcan1/2 deficient MEFs. Functionally, TAK1 was indispensable for the cardiomyocyte growth response induced by pro-hypertrophic stimuli through calcineurin. These results describe a signalling relationship between two central regulatory pathways in which TAK1–TAB1–TAB2 selectively induces calcineurin–NFAT signalling through direct phosphorylation of RCAN1, while calcineurin activation diminishes TAK1 signalling by dephosphorylation of TAK1 and TAB1.

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Figure 1: RCAN1 physically interacts with TAB2.
Figure 2: TAK1 phosphorylates RCAN1 at Ser 94 and 136.
Figure 3: TAK1 induces calcineurin–NFAT signalling in cardiomyocytes.
Figure 4: Calcineurin–RCAN1 is required for TAK1-induced hypertrophic growth in cardiomyocytes.
Figure 5: Calcineurin reciprocally regulates TAK1 activity.

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Acknowledgements

We are very grateful to Yang Xia for the generous gift of the S108/112A mutant RCAN1 plasmid, and Jun Tsuji for providing the Tak1−/− MEFs originated from Akira's laboratory. This work was supported by grants from the National Institutes of Health (J.D.M), the Fondation Leducq (Heart failure network grant to J.D.M), and the Howard Hughes Medical Institute.

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Authors and Affiliations

  1. Department of Pediatrics, University of Cincinnati, Cincinnati Children's Hospital Medical Center, and the Howard Hughes Medical Institute, Cincinnati, 45229, Ohio, USA

    Qinghang Liu & Jeffery D. Molkentin

  2. The Scripps Research Institute, Jupiter, 33458, Florida, USA

    Jennifer Caldwell Busby

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  1. Qinghang Liu
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  2. Jennifer Caldwell Busby
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  3. Jeffery D. Molkentin
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Contributions

Q.L. performed the experiments; J.O.M. supervised all work and data interpretation; J.C.B. performed mass spectrometry of phosphorylated RCAN1.

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Correspondence to Jeffery D. Molkentin.

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The authors declare no competing financial interests.

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Liu, Q., Busby, J. & Molkentin, J. Interaction between TAK1–TAB1–TAB2 and RCAN1–calcineurin defines a signalling nodal control point. Nat Cell Biol 11, 154–161 (2009). https://doi.org/10.1038/ncb1823

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