title: Bone morphogenetic protein-2 induces expression of murine zinc finger transcription factor ZNF450.
abstract: The bone morphogenetic protein-2 (BMP-2) is a potent secreted factor that promotes osteoblast differentiation during development. Exposure to BMP-2 is sufficient to cause a lasting change in cell fate presumably by activating specific target genes. To identify genes downstream of BMP-2 we treated the murine pluripotent embryonic cell line, C3H10T1/2 that can be induced to form an osteoblastic phenotype, with 100 ng/ml BMP-2 for 24 h. Using suppression subtractive hybridisation we found the novel zinc finger transcription factor, ZNF450 was upregulated. The single-copy ZNF450 gene spans 15.6 kb on chromosome 10B1 and consists of seven exons, the first of which is untranslated. The open reading frame encodes a 710 reside protein. Analysis of the protein sequence reveals a highly conserved amino-terminal BTB/POZ dimerisation domain, an AT-hook motif, and eight C2H2 zinc fingers. Library screening identified a second mRNA isoform encoding a short protein isoform with one zinc finger. Using reverse transcriptase-real time PCR to measure mRNA expression we found that ZNF450, Runx2/Cbfa-1, and Sp7/osterix were induced by BMP-2 after 4 h in C2C12 myoblast cells. Treatment of C2C12 cells with BMP-2 causes a shift from a myoblastic to osteoblastic phenotype. ZNF450 was upregulated three to fivefold after 24 h in C3H10T1/2 cells and required 100 ng/ml BMP-2. Expression of the 3 kb major transcript was highest in liver, testis, and kidney. However, ZNF450 mRNA was found also in a wide range of adult tissues. The consistent induction of ZNF450 by BMP-2 after 4 h in three murine pluripotent cell lines suggests that ZNF450 may play a role in the BMP-2 signalling pathway.
authors.affiliation: Department of Craniofacial Development, GKT Dental Institute, King's College, Guy's Hospital, London SE1 9RT, United Kingdom. alasdair.edgar@kcl.ac.uk
authors.forename: Alasdair J
authors.initials: AJ
authors.lastname: Edgar
authors.forename: Sharon L
authors.initials: SL
authors.lastname: Dover
authors.forename: Melanie N
authors.initials: MN
authors.lastname: Lodrick
authors.forename: Ian J
authors.initials: IJ
authors.lastname: McKay
authors.forename: Francis J
authors.initials: FJ
authors.lastname: Hughes
authors.forename: Wendy
authors.initials: W
authors.lastname: Turner
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D015152:Blotting, Northern; D055396:Bone Morphogenetic Protein 2; D019485:Bone Morphogenetic Proteins / Q000502:physiology*; D002460:Cell Line; D051379:Mice; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D013382:Subtraction Technique; D014157:Transcription Factors / Q000737:chemistry / Q000502:physiology*; D016212:Transforming Growth Factor beta / Q000502:physiology*; D016335:Zinc Fingers
chemical_list: C525719:Bmp2 protein, mouse; D055396:Bone Morphogenetic Protein 2; D019485:Bone Morphogenetic Proteins; D012333:RNA, Messenger; D014157:Transcription Factors; D016212:Transforming Growth Factor beta; C501496:Zbtb24 protein, mouse