Regulation of chondrocyte differentiation by IRE1α depends on its enzymatic activity

Bone morphogenetic protein 2(BMP2) is known to activate unfolded protein response (UPR) signal molecules in chondrogenesis. Inositol-requiring enzyme-1 alpha (IRE1 alpha),as one of three unfolded protein sensors in UPR signaling pathways, can be activated during ER stress. However, the influence on IRE1 alpha in chondrocyte differentiation has not yet been elucidated. Here we present evidence demonstrating that overexpression of IRE1 alpha inhibits chondrocyte differentiation, as revealed by reduced expression of collagen II (Corn), Sox9, collagen X (CoIX), matrix metalloproteinase 13 (MMP-13), Indian hedgehog (IHH), Runx2 and enhanced expression of parathyroid hormone-related peptide (PTHrP). Furthermore, IRE1 alpha-mediated inhibition of chondrogenesis depends on its enzymatic activity, since its point mutant lacking enzymatic activity completely loses this activity. The RNase and Kinase domains of IRE1 alpha C-terminal are necessary for its full enzymatic activity and inhibition of chondrocyte differentiation. Mechanism studies demonstrate that granulin-epithelin precursor(GEP), a growth factor known to stimulate chondrogenesis, induced IRE1 alpha expression in chondrogenesis. The expression of IRE1 alpha is depended on GEP signaling, and IRE1ot expression is hardly detectable in GEP(-/-) embryos. In addition, IRE1 alpha inhibits GEP-mediated chondrocyte differentiation as a negative regulator. Altered expression of IRE1 alpha in chondrocyte hypertrophy was accompanied by altered levels of IHH and PTHrP. Collectively, IRE1 alpha may be a novel regulator of chondrocyte differentiation by 1) inhibition GEP-mediated chondrocyte differentiation as a negative regulator; 2) promoting IHH/PTHrP signaling. (C) 2014 Elsevier Inc. All rights reserved.