Abstract
Macrophages play pivotal roles in development, homeostasis, tissue repair and immunity. Macrophage proliferation is promoted by macrophage colony-stimulating factor (M-CSF)-induced Akt signaling; yet, how this process is terminated remains unclear. Here, we identify casein kinase 2-interacting protein-1 (CKIP-1) as a novel inhibitor of macrophage proliferation. In resting macrophages, CKIP-1 was phosphorylated at Serine 342 by constitutively active GSK3β, the downstream target of Akt. This phosphorylation triggers the polyubiquitination and proteasomal degradation of CKIP-1. Upon M-CSF stimulation, Akt is activated by CSF-1R-PI3K and then inactivates GSK3β, leading to the stabilization of CKIP-1 and β-catenin proteins. β-catenin promotes the expression of proliferation genes including cyclin D and c-Myc. CKIP-1 interacts with TRAF6, a ubiquitin ligase required for K63-linked ubiquitination and plasma membrane recruitment of Akt, and terminates TRAF6-mediated Akt activation. By this means, CKIP-1 inhibits macrophage proliferation specifically at the late stage after M-CSF stimulation. Furthermore, CKIP-1 deficiency results in increased proliferation and decreased apoptosis of macrophages in vitro and CKIP-1−/− mice spontaneously develop a macrophage-dominated splenomegaly and myeloproliferation. Together, these data demonstrate that CKIP-1 plays a critical role in the regulation of macrophage homeostasis by inhibiting TRAF6-mediated Akt activation.
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Acknowledgements
We thank Drs Jiang Peng for RAW264.7 cell line, Changyan Li for lentiviral transduction, Hongbing Shu for TARF6 plasmid, Jiyan Zhang for BMDM cultures. This work was supported by Chinese National Basic Research Programs (2011CB910802, 2012CB910304), the National Key Technologies R&D Program for New Drugs (2014ZX09J14106-04C), Chinese National Natural Science Foundation Projects (31125010, 31270912 and 81221004) and a grant from State Key Laboratory of Proteomics (SKLP-K201102).
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( Supplementary information is linked to the online version of the paper on the Cell Research website.)
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Supplementary information, Figure S1
Normal differentiation of CKIP-1−/− BMDMs. (PDF 156 kb)
Supplementary information, Figure S2
CKIP-1 silencing augments M-CSF-induced Akt activation. (PDF 264 kb)
Supplementary information, Figure S3
TRAF6 is required for M-CSF-induced Akt activation. (PDF 307 kb)
Supplementary information, Figure S4
PI3K/Akt/GSK3β pathway controls CKIP-1 protein level in macrophage. (PDF 326 kb)
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Zhang, L., Wang, Y., Xiao, F. et al. CKIP-1 regulates macrophage proliferation by inhibiting TRAF6-mediated Akt activation. Cell Res 24, 742–761 (2014). https://doi.org/10.1038/cr.2014.53
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DOI: https://doi.org/10.1038/cr.2014.53
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