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Calpain Modulates Cyclin-Dependent Kinase Inhibitor 1B (p27Kip1) in Cells of the Osteoblast Lineage

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Abstract

The ubiquitously expressed calpains-1 and -2 belong to a family of calcium-dependent intracellular cysteine proteases. Both calpains are heterodimers consisting of a large catalytic subunit and a small regulatory subunit encoded by the gene Capn4. Ablation of the calpain small subunit eliminates calpain activity and leads to embryonic lethality. We previously created osteoblast-specific Capn4 knockout mice to investigate a physiological role for the calpain small subunit in cells of the osteoblast lineage. Deletion of Capn4 reduced trabecular and cortical bone, mainly due to impaired proliferation and differentiation of cells of the osteoblast lineage. To further investigate an underlining mechanism by which osteoblast-specific Capn4 knockout mice develop an osteoporotic bone phenotype, we established osteoblastic cell lines stably expressing either control or Capn4 RNA interference for this study. Capn4 knockdown cells showed reduced cell proliferation, accumulation of total and phosphorylated cyclin-dependent kinase inhibitor 1B (p27Kip1) on serine 10, and reduced phosphorylation of retinoblastoma protein on threonine 821. Moreover, ablation of Capn4 increased 27 Kip1 mRNA levels, likely due to stabilized binding of Akt to protein phosphatase 2A, which presumably results in reduced phosphorylation of Akt on S473 and forkhead Box O (FoxO) 3A on T32. Collectively, calpain regulates cell proliferative function by modulating both transcription and degradation of p27Kip1 in osteoblasts. In conclusion, calpain is a critical modulator for regulation of p27Kip1 in cells of the osteoblast lineage.

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Acknowledgements

We thank Dr. Hanno Hock and Laura B. Prickett-Rice at the Center for Regenerative Medicine, Massachusetts General Hospital (MGH), for consultation and assistance with the flow-cytometric analysis. This work was partially supported by National Institutes of Health grant R01 DK072102, the William F. Milton Fund, and the MGH interim support fund (to M. S.).

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Correspondence to Masako Shimada.

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The authors have stated that they have no conflict of interest.

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Kashiwagi, A., Fein, M.J. & Shimada, M. Calpain Modulates Cyclin-Dependent Kinase Inhibitor 1B (p27Kip1) in Cells of the Osteoblast Lineage. Calcif Tissue Int 89, 36–42 (2011). https://doi.org/10.1007/s00223-011-9491-3

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