title: Transcriptional regulation of bone sialoprotein gene by interleukin-11.
abstract: Interleukin-11 (IL-11) is a stromal cell-derived cytokine that belongs to the interleukin-6 family of cytokines. IL-11 has many biological activities and has roles in hematopoiesis, immune responses, the nervous system and bone metabolism. Bone sialoprotein (BSP) is a mineralized tissue-specific protein expressed in differentiated osteoblasts that appears to function in the initial mineralization of bone. IL-11 (20 ng/ml) increased BSP mRNA and protein levels at 12h in osteoblast-like ROS 17/2.8 cells. In a transient transfection assay, IL-11 (20 ng/ml) increased luciferase activity of the construct (-116 to +60) in ROS 17/2.8 cells and rat bone marrow stromal cells. Introduction of 2 bp mutations to the luciferase constructs showed that the effects of IL-11 were mediated by a cAMP response element (CRE), a fibroblast growth factor 2 response element (FRE) and a homeodomain protein-binding site (HOX). Luciferase activities induced by IL-11 were blocked by protein kinase A inhibitor, tyrosine kinase inhibitor and ERK1/2 inhibitor. Gel shift analyses showed that IL-11 (20 ng/ml) increased nuclear protein binding to CRE, FRE and HOX. CREB1, phospho-CREB1, c-Fos, c-Jun, JunD and Fra2 antibodies disrupted the formation of CRE-protein complexes. Dlx5, Msx2, Runx2 and Smad1 antibodies disrupted FRE- and HOX-protein complex formations. These studies demonstrate that IL-11 stimulates BSP transcription by targeting CRE, FRE and HOX sites in the proximal promoter of the rat BSP gene. Moreover, phospho-CREB1, c-Fos, c-Jun, JunD, Fra2, Dlx5, Msx2, Runx2 and Smadl transcription factors appear to be key regulators of IL-11 effects on BSP transcription.
authors.affiliation: Department of Periodontology, Nihon University School of Dentistry at Matsudo, Chiba, 271-8587, Japan.
authors.forename: Shuang
authors.initials: S
authors.lastname: Wang
authors.forename: Yoko
authors.initials: Y
authors.lastname: Sasaki
authors.forename: Liming
authors.initials: L
authors.lastname: Zhou
authors.forename: Hiroyoshi
authors.initials: H
authors.lastname: Matsumura
authors.forename: Shouta
authors.initials: S
authors.lastname: Araki
authors.forename: Masaru
authors.initials: M
authors.lastname: Mezawa
authors.forename: Hideki
authors.initials: H
authors.lastname: Takai
authors.forename: Zhen
authors.initials: Z
authors.lastname: Chen
authors.forename: Yorimasa
authors.initials: Y
authors.lastname: Ogata
mesh_terms: D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D050659:Core Binding Factor Alpha 1 Subunit / Q000235:genetics; D017931:DNA Primers / Q000235:genetics; D058574:Integrin-Binding Sialoprotein / Q000235:genetics* / Q000378:metabolism; D017370:Interleukin-11 / Q000494:pharmacology*; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D010006:Osteoblasts / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D051381:Rats; D011994:Recombinant Proteins / Q000494:pharmacology; D014157:Transcription Factors / Q000235:genetics; D015533:Transcriptional Activation / Q000187:drug effects; D014162:Transfection
chemical_list: D050659:Core Binding Factor Alpha 1 Subunit; D017931:DNA Primers; D058574:Integrin-Binding Sialoprotein; D017370:Interleukin-11; D012333:RNA, Messenger; D011994:Recombinant Proteins; C493717:Runx2 protein, rat; C495181:Sp7 protein, rat; D014157:Transcription Factors