title: Dual involvement of protein kinase C delta in apoptosis induced by syndecan-2 in osteoblasts.
abstract: Syndecans are proteoglycans that act as signaling molecules. Previously, we showed that syndecan-2 (SYND2) is involved in the control of osteoblastic (OB) cell apoptosis. Here, we show a novel functional interaction between SYND2 and protein kinase C delta (PKCdelta). Overexpression of SYND2 in MG63 OB cells resulted in increased PKCdelta protein level without change in PKCdelta mRNA production. In SYND2-transfected cells, the increase in PKCdelta was restricted to the cytosolic compartment, threonine 505-PKCdelta was underphosphorylated and immunoprecipitated PKCdelta showed decreased capacity to phosphorylate histone, indicating that SYND2 decreased PKCdelta activity. Inhibition of PKCdelta by Rottlerin or a dead-kinase dominant negative (DN) construct activated effector caspases and increased the number of apoptotic cells. In addition, rescue of kinase activity with a construct coding, the PKCdelta catalytic domain (CAT) reduced SYND2-induced apoptosis. This indicates that PKCdelta acts as a pro-survival kinase and that SYND2 inhibits the anti-apoptotic action of PKCdelta in OB cells. We also showed that overexpression of PKCdelta wild type (WT) induced osteoblast apoptosis. Moreover, inhibition of PKCdelta by siRNA resulted in increased apoptosis in control cells but reduced apoptosis in SYND2-overexpressing osteoblasts, indicating that SYND2 requires PKCdelta accumulation to induce apoptosis. These results show that SYND2 modulates PKCdelta actions by inhibition of the canonical allosterical activation pathway that plays an anti-apoptotic role in OB cells, and promotion of a pro-apoptotic role that may depend on PKCdelta protein level and that participates to the induction of cell death by SYND2. This establishes a functional interaction between SYND2 and PKCdelta in osteoblasts.
authors.affiliation: Laboratory of Osteoblast Biology and Pathology, INSERM, Lariboisière Hospital, University Paris 7, Paris, France.
authors.forename: Armelle
authors.initials: A
authors.lastname: Orosco
authors.forename: Olivia
authors.initials: O
authors.lastname: Fromigué
authors.forename: Eric
authors.initials: E
authors.lastname: Haÿ
authors.forename: Pierre J
authors.initials: PJ
authors.lastname: Marie
authors.forename: Dominique
authors.initials: D
authors.lastname: Modrowski
mesh_terms: D017209:Apoptosis / Q000502:physiology*; D002460:Cell Line; D002470:Cell Survival / Q000502:physiology; D006801:Humans; D008562:Membrane Glycoproteins / Q000235:genetics / Q000378:metabolism*; D010006:Osteoblasts / Q000378:metabolism*; D010766:Phosphorylation; D051745:Protein Kinase C-delta / Q000037:antagonists & inhibitors / Q000378:metabolism*; D011499:Protein Processing, Post-Translational / Q000502:physiology; D011509:Proteoglycans / Q000235:genetics / Q000378:metabolism*; D015398:Signal Transduction / Q000502:physiology*; D053669:Syndecan-2; D014162:Transfection
chemical_list: D008562:Membrane Glycoproteins; D011509:Proteoglycans; C508245:SDC2 protein, human; D053669:Syndecan-2; D051745:Protein Kinase C-delta